Proceedings Volume 10024

Optics in Health Care and Biomedical Optics VII

Qingming Luo, Xingde Li, Ying Gu, et al.
Proceedings Volume 10024

Optics in Health Care and Biomedical Optics VII

Qingming Luo, Xingde Li, Ying Gu, et al.
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Volume Details

Date Published: 23 December 2016
Contents: 9 Sessions, 100 Papers, 0 Presentations
Conference: SPIE/COS Photonics Asia 2016
Volume Number: 10024

Table of Contents


Table of Contents

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  • Front Matter: Volume 10024
  • Advanced Optical Techniques for Clinical Medicine II
  • Multimodal Biomedical Imaging
  • Tissue Optics
  • Microscopy and Imaging II
  • Nano/Biophotonics
  • Photon Therapeutics
  • Biomedical Spectroscopy
  • Poster Session
Front Matter: Volume 10024
Front Matter: Volume 10024
This PDF file contains the front matter associated with SPIE Proceedings Volume 10024, including the Title Page, Copyright information, Table of Contents, and Conference Committee listing.
Advanced Optical Techniques for Clinical Medicine II
Detection and classification of ebola on microfluidic chips
Xue Lin, Xiangyu Jin, Yunqian Fan, et al.
Point-of-care testing (POCT) for an infectious diseases is the prerequisite to control of the disease and limitation of its spread. A microfluidic chip for detection and classification of four strains of Ebola virus was developed and evaluated. This assay was based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) and specific primers for Ebola Zaire virus, Ebola Sudan virus, Ebola Tai Forest virus and Ebola Bundibugyo virus were designed. The sensitivity of the microfluidic chip was under 103 copies per milliliter, as determined by ten repeated tests. This assay is unique in its ability to enable diagnosis of the Ebola infections and simultaneous typing of Ebola virus on a single chip. It offers short reaction time, ease of use and high specificity. These features should enable POCT in remote area during outbreaks of Ebola virus.
Multimodal Biomedical Imaging
Studying the numeration methods of signals with unstable background for in vivo flow cytometry
Xiaoling Wang, Yuanzhen Suo, Dan Wei, et al.
In recent years, the in vivo flow cytometry (IVFC) has been a useful technology in detecting and quantifying the circulating cells dynamically in living animals, especially in the research related to the cell tracking and the cancer metastasis. In practice, however, the unstable background signals caused by the experiment animals’ respiratory movement, limb movement and photo-bleaching of tissues’ auto-fluorescence exist in many IVFC data, which could affect the accuracy of cell counting results in the following post-processing procedure, making the IVFC signals less available. Here we developed a signal processing method that could effectively correct the unstable background signals by using methods combining interpolating, fitting, automatic segmenting and wavelet-based denoising. Compared with the previously used non-correction methods, i.e., the “line-gating” method or the automatic threshold method, this method showed a higher accuracy and efficiency in counting cell numbers of IVFC signals, as well as demonstrating a better statistic results in the Pearson’s correlation coefficient R2 and the mean-squared error (MSE).
Microwave-induced thermoacoustic imaging system based on flexible transducer
Microwave-induced thermoacoustic (TA) imaging combines the advantages of high imaging contrast due to electromagnetic absorption and high resolution of the ultrasound technology, and it is a potential alternative imaging technique for biomedical applications, particularly for breast tumor detection. The traditional TA system uses circular-scanning (CS) to obtain complete information to reconstruct a two-dimensional image, however, it needs a large operating space for rotation of the transducers and bulk of coupling medium limiting its medical applications. The linear-scanning (LS) system can overcome these problems partially but usually lose some information and cause image distortion. In this paper, in order to overcome above limitations, a TA imaging system with Sample-Cling-Scanning (SCS) model based on a flexible multi-element transducer is presented. It combines the advantages of both CS and LS modes, and overcome their limitations. Meanwhile, an adaptive back projection algorithm is presented to implement this scanning model. The experimental results show that the proposed system combines advantages including shape adaptation, information integrity, and efficient transmission. These advantages make it a preferred system for TA applications, especially in breast tumor detection.
All-optically integrated multimodality imaging system: combined photoacoustic microscopy, optical coherence tomography, and fluorescence imaging
We have developed a multimodality imaging system by optically integrating all-optical photoacoustic microscopy (AOPAM), optical coherence tomography (OCT) and fluorescence microscopy (FLM) to provide complementary information including optical absorption, optical back-scattering and fluorescence contrast of biological tissue. By sharing the same low-coherence Michelson interferometer, AOPAM and OCT could be organically optically combined to obtain the absorption and scattering information of the biological tissues. Also, owing to using the same laser source and objective lens, intrinsically registered photoacoustic and fluorescence signals are obtained to present the radiative and nonradiative transition process of absorption. Simultaneously photoacoustic angiography, tissue structure and fluorescence molecular in vivo images of mouse ear were acquired to demonstrate the capabilities of the optically integrated trimodality imaging system, which can present more information to study tumor angiogenesis, vasculature, anatomical structure and microenvironments in vivo.
Extinction measurement of dense media by an optical coherence tomography technique
Tomoki Ago, Toshiaki Iwai, Ryoko Yokota
The optical coherence tomography will make progress as the next stage toward a spectroscopic analysis technique. The spectroscopic analysis is based on the Beer-Lambert law. The absorption and scattering coefficients even for the dense medium can be measured by the Beer-Lambert law because the OCT can detect only the light keeping the coherency which propagated rectilinearly and retro-reflected from scatters. This study is concerned with the quantitative verification of Beer-Lambert law in the OCT imaging.
Tissue Optics
Microfluidic cytometers with integrated on-chip optical components for blood cell analysis
In the last two decades, microfluidic technologies have shown the great potential in developing portable and point-of care testing blood cell analysis devices. It is challenging to integrate all free-space detecting components in a single microfluidic platform. In this paper, a microfluidic cytometer with integrated on-chip optical components was demonstrated. To facilitate on-chip detection, the device integrated optical fibers and on-chip microlens with microfluidic channels on one polydimethylsiloxane layer by standard soft photolithography. This compact design increased the sensitivity of the device and also eliminated time-consuming free-space optical alignments. Polystyrene particles, together with red blood cells and platelets, were measured in the microfluidic cytometer by small angle forward scatter. Experimental results indicated that the performance of the microfluidic device was comparable to a conventional cytometer. And it was also demonstrated its ability to detect on-chip optical signals in a highly compact, simple, truly portable and low cost format which was perfect suitable for point-of-care testing clinical hematology diagnostics.
Weighted optimization of irradiance for photodynamic therapy of port wine stains
Planning of irradiance distribution (PID) is one of the foremost factors for on-demand treatment of port wine stains (PWS) with photodynamic therapy (PDT). A weighted optimization method for PID was proposed according to the grading of PWS with a three dimensional digital illumination instrument. Firstly, the point clouds of lesions were filtered to remove the error or redundant points, the triangulation was carried out and the lesion was divided into small triangular patches. Secondly, the parameters such as area, normal vector and orthocenter for optimization of each triangular patch were calculated, and the weighted coefficients were determined by the erythema indexes and areas of patches. Then, the optimization initial point was calculated based on the normal vectors and orthocenters to optimize the light direction. In the end, the irradiation can be optimized according to cosine values of irradiance angles and weighted coefficients. Comparing the irradiance distribution before and after optimization, the proposed weighted optimization method can make the irradiance distribution match better with the characteristics of lesions, and has the potential to improve the therapeutic efficacy.
Stage scoring of liver fibrosis using Mueller matrix microscope
Jialing Zhou, Honghui He, Ye Wang, et al.
Liver fibrosis is a common pathological process of varied chronic liver diseases including alcoholic hepatitis, virus hepatitis, and so on. Accurate evaluation of liver fibrosis is necessary for effective therapy and a five-stage grading system was developed. Currently, experienced pathologists use stained liver biopsies to assess the degree of liver fibrosis. But it is difficult to obtain highly reproducible results because of huge discrepancy among different observers. Polarization imaging technique has the potential of scoring liver fibrosis since it is capable of probing the structural and optical properties of samples. Considering that the Mueller matrix measurement can provide comprehensive microstructural information of the tissues, in this paper, we apply the Mueller matrix microscope to human liver fibrosis slices in different fibrosis stages. We extract the valid regions and adopt the Mueller matrix polar decomposition (MMPD) and Mueller matrix transformation (MMT) parameters for quantitative analysis. We also use the Monte Carlo simulation to analyze the relationship between the microscopic Mueller matrix parameters and the characteristic structural changes during the fibrosis process. The experimental and Monte Carlo simulated results show good consistency. We get a positive correlation between the parameters and the stage of liver fibrosis. The results presented in this paper indicate that the Mueller matrix microscope can provide additional information for the detections and fibrosis scorings of liver tissues and has great potential in liver fibrosis diagnosis.
3D printing of tissue-simulating phantoms for calibration of biomedical optical devices
Zuhua Zhao, Ximing Zhou, Shuwei Shen, et al.
Clinical utility of many biomedical optical devices is limited by the lack of effective and traceable calibration methods. Optical phantoms that simulate biological tissues used for optical device calibration have been explored. However, these phantoms can hardly simulate both structural and optical properties of multi-layered biological tissue. To address this limitation, we develop a 3D printing production line that integrates spin coating, light-cured 3D printing and Fused Deposition Modeling (FDM) for freeform fabrication of optical phantoms with mechanical and optical heterogeneities. With the gel wax Polydimethylsiloxane (PDMS), and colorless light-curable ink as matrix materials, titanium dioxide (TiO2) powder as the scattering ingredient, graphite powder and black carbon as the absorption ingredient, a multilayer phantom with high-precision is fabricated. The absorption and scattering coefficients of each layer are measured by a double integrating sphere system. The results demonstrate that the system has the potential to fabricate reliable tissue-simulating phantoms to calibrate optical imaging devices.
Effect of surface topographic features on the optical properties of skin: a phantom study
Guangli Liu, Jianfeng Chen, Zuhua Zhao, et al.
Tissue-simulating phantoms are used to validate and calibrate optical imaging systems and to understand light transport in biological tissue. Light propagation in a strongly turbid medium such as skin tissue experiences multiple scattering and diffuse reflection from the surface. Surface roughness introduces phase shifts and optical path length differences for light which is scattered within the skin tissue and reflected from the surface. In this paper, we study the effect of mismatched surface roughness on optical measurement and subsequent determination of optical properties of skin tissue. A series of phantoms with controlled surface features and optical properties corresponding to normal human skin are fabricated. The fabrication of polydimethylsiloxane (PDMS) phantoms with known surface roughness follows a standard soft lithography process. Surface roughness of skin-simulating phantoms are measured with Bruker stylus profiler. The diffuse reflectance of the phantom is validated by a UV/VIS spectrophotometer. The results show that surface texture and roughness have considerable influence on the optical characteristics of skin. This study suggests that surface roughness should be considered as an important contributing factor for the determination of tissue optical properties.
Structured light imaging system for structural and optical characterization of 3D tissue-simulating phantoms
Songde Liu, Zach Smith, Ronald X. Xu
There is a pressing need for a phantom standard to calibrate medical optical devices. However, 3D printing of tissue-simulating phantom standard is challenged by lacking of appropriate methods to characterize and reproduce surface topography and optical properties accurately. We have developed a structured light imaging system to characterize surface topography and optical properties (absorption coefficient and reduced scattering coefficient) of 3D tissue-simulating phantoms. The system consisted of a hyperspectral light source, a digital light projector (DLP), a CMOS camera, two polarizers, a rotational stage, a translation stage, a motion controller, and a personal computer. Tissue-simulating phantoms with different structural and optical properties were characterized by the proposed imaging system and validated by a standard integrating sphere system. The experimental results showed that the proposed system was able to achieve pixel-level optical properties with a percentage error of less than 11% for absorption coefficient and less than 7% for reduced scattering coefficient for phantoms without surface curvature. In the meanwhile, 3D topographic profile of the phantom can be effectively reconstructed with an accuracy of less than 1% deviation error. Our study demonstrated that the proposed structured light imaging system has the potential to characterize structural profile and optical properties of 3D tissue-simulating phantoms.
Microscopy and Imaging II
A frequency domain reconstruction of SIM image using four raw images
This work is a result of an exploration in response to the following observation: though reconstruction methodology of SIM image since its origin is carried out in frequency domain, recent works in this field, specially when it comes to SIM reconstruction using lesser than 9 raw images, have ventured into reconstruction methodologies that operate directly in the spatial domain. This work formulates and demonstrates a frequency domain reconstruction of SIM image using four raw images – one wide-field image and three SIM images. One of the chief feature of the presented reconstruction algorithm is that it employs the standard ‘tools and tricks’ used by the conventional 9-frame SIM reconstruction algorithm. Results indicate that the presented reconstruction algorithm provides high resolution reconstructions successfully as long as the noise level in the raw images is lower than 10%. For higher level of noise, the reconstruction result shows little resolution enhancement.
Multiple gold-dimer detection from large scattering background
Xin Hong, Zheng Jin
Gold nanoparticles exhibit unique plasmonic optical properties in visible to near infrared band. Especially the coupling effect existing at the gap between a closely linked particle pair can make the local field strongly enhanced. These properties make gold particles more attractive to be employed as molecular probes in biomedical related fundamental and clinical researches. However in the bio-system exist many large molecules or groups, whose optical signals can strongly depress the gold particles without detectable. In this paper, we proposed a method to extract the targets which are labelled by gold dimer pairs from large scattering background.
Anti-hepatocarcinoma effects of berberine-nanostructured lipid carriers against human HepG2, Huh7, and EC9706 cancer cell lines
Xiang-Ping Meng, Hua Fan, Yi-fei Wang, et al.
Hepatocarcinoma and esophageal squamous cell carcinomas threaten human life badly. It is a current issue to seek the effective natural remedy from plant to treat cancer due to the resistance of the advanced hepatocarcinoma and esophageal carcinoma to chemotherapy. Berberine (Ber), an isoquinoline derivative alkaloid, has a wide range of pharmacological properties and is considered to have anti-hepatocarcinoma and antiesophageal carcinoma effects. However its low oral bioavailability restricts its wide application. In this report, Ber loaded nanostructured lipid carriers (Ber-NLC) was prepared by hot melting and then high pressure homogenization technique. The in vitro anti-hepatocarcinoma and antiesophageal carcinoma effects of Ber-NLC relative to efficacy of bulk Ber were evaluated. The particle size and zeta potential of Ber-NLC were 189.3 ± 3.7 nm and −19.3 ± 1.4 mV, respectively. MTT assay showed that Ber-NLC effectively inhibited the proliferation of human HepG2 and Huh7 and EC9706 cells, and the corresponding IC50 value was 9.1 μg/ml, 4.4 μg/ml, and 6.3 μg/ml (18.3μg/ml, 6.5μg/ml, and 12.4μg/ml μg/ml of bulk Ber solution), respectively. These results suggest that the delivery of Ber-NLC is a promising approach for treating tumors.
Evaluation of free radical scavenging and anti-oxidative capacity of polydatin-nanostructured lipid carriers
Xiang-Ping Meng, Fan Shi, Hai-Jie Li, et al.
Cellular damage induced by free-radicals like reactive oxygen species has been implicated in several diseases. 2, 2-azobis(2-amidino-propane) dihydrochloride(AAPH) generates two potent ROS capable of inducing lipid peroxidation: alkoxy radical(RO-) and peroxy radical (ROO-). These radicals are similar to those that are physiologically active and thus might initiate a cascade of intracellular toxic events leading to oxidation, lipid peroxidation, DNA damage and subsequent cell death. Hence naturally anti-oxidant play a vital role in combating these conditions. In this study, polydatin loaded nanostructured lipid carriers (Pol-NLC) was prepared by hot melting and then high pressure homogenization technique. The effects of Pol-NLC on free radical scavenging and anti-oxidative capacity is investigated. The particle size and zeta potential of Pol-NLC were 113.9 ± 1.1 nm and −16.3 1 ± 0.27 mV, respectively. By free radical scavenging assays, the IC50 value of Pol-NLC were 28.71, 9.83 μg/mL with DPPH, ABTS assay respectively, and 0.143 mg ferrous sulfate/1 mg Pol-NLC with FRAP assay. These results indicated that the antioxidant properties of Pol-NLC hold great potential used as an alternative to more toxic synthetic anti-oxidants as an additive in food, cosmetic and pharmaceutical preparations for the oxidative diseases treatment.
Photon Therapeutics
Quantification of reactive oxygen species for photodynamic therapy
Zou Tan, Jinde Zhang, Lisheng Lin, et al.
Photodynamic therapy (PDT) is an effective therapeutic modality that uses a light source to activate light-sensitive photosensitizers to treat both oncologic and nononcological indications. Photosensitizers are excited to the long-lived triplet state, and they react with biomolecules via type I or II mechanism resulted in cell death and tumor necrosis. Free radicals and radical ions are formed by electron transfer reactions (type I), which rapidly react with oxygen leading to the production of reactive oxygen species (ROS), including superoxide ions, hydroxyl radicals and hydrogen peroxide. Singlet molecular oxygen is produced in a Type II reaction, in which the excited singlet state of the photosensitizer generated upon photon absorption by the ground-state photosensitizer molecule undergoes intersystem crossing to a long-lived triplet state. In this talk, the fundmental mechanisms and detection techniques for ROS generation in PDT will be introduced. In particular, the quantification of singlet oxygen generation for pre-clinical application will be highlighted, which plays an essential role in the establishment of robust singlet oxygen-mediated PDT dosimetry.
In vitro photodynamic inactivation effects of cationic benzylidene cyclopentanone photosensitizers on clinical fluconazole-resistant Candida albicans planktonic cells and biofilms
Shaona Zhou, Yanyan Fang, Zulin Ye, et al.
Background: An increasing prevalence of Candida infections has emerged with the wide use of immune-suppressants and antibiotics. Photodynamic inactivation (PDI) as a new approach to treat localized Candida infections is an emerging and promising field nowadays. This study evaluated the efficacy of photodynamic therapy using two new Cationic benzylidene cyclopentanone photosensitizers(P1 and P2) against strains of clinical fluconazole–resistant Candida albicans.

Methods: Suspensions and biofilms of Candida species were incubated with P1 and P2 concentrations (0.25~50 μM) for 30 min followed by 532nm laser irradiation. For planktonic suspensions, viability of cells was assayed by CFU counting. For biofilms, the metabolic activity was evaluated by XTT. Results: In PDI of a planktonic culture of clinical fluconazole–resistant Candida albicans, P2 showed the higher efficacy. After incubation with 25 μM of P2 for 30 min and irradiation with 532nm laser (36 J cm-2), the viability of C. albicans planktonic cells decreased by 3.84 log10. For biofilm cells, a higher light dose of 75 mW cm-2 was necessary to achieve 97.71% metabolic activity reduction.

Conclusions: The results of this investigation demonstrated that benzylidene cyclopentanone photosensitizer(P2)is an efficient photosensitizer to kill C. albicans. Moreover, single-species biofilms were less susceptible to PDT than their planktonic counterparts.
Antimicrobial blue light inactivation of Methicillin-resistant Staphylococcus aureus
Background: With the increasing emergence of multidrug-resistant (MDR) bacterial strains, there is a pressing need for the development of alternative treatment for infections. Antimicrobial blue light (aBL) has provided a simple and effective approach.

Methods: We first investigated the effectiveness of aBL (415 nm) inactivation of USA300 LAClux (a communityacquired Methicillin-resistant Staphylococcus aureus strain) both in the planktonic and biofilm forms. The survival of the bacteria in suspensions was determined by serial dilution and that of the biofilm-embedded bacteria was determined by bioluminescence quantification. Using a mouse model of thermal burn infected with USA300 LAClux, we further assessed the effectiveness of aBL for treating localized infections. Bioluminescence imaging was performed to monitor in real time bacterial viability in vivo.

Results: In vitro study showed that, for the planktonic counterpart of the bacteria or the 24-h-old biofilms, an irradiance of 55 mW/cm2 for 60 min resulted in a 4.61 log10 or 2.56 log10 inactivation, respectively. In vivo study using infected mouse burns demonstrated that a 2.56-log10 inactivation was achieved after 100-mW/cm2 irradiation for 62 min. Conclusions: aBL is a potential alternative approach for treating Methicillin-resistant Staphylococcus aureus infections.
Biomedical Spectroscopy
Quantitatively differentiating microstructural variations of skeletal muscle tissues by multispectral Mueller matrix imaging
Polarized light is sensitive to the microstructures of biological tissues and can be used to detect physiological changes. Meanwhile, spectral features of the scattered light can also provide abundant microstructural information of tissues. In this paper, we take the backscattering polarization Mueller matrix images of bovine skeletal muscle tissues during the 24-hour experimental time, and analyze their multispectral behavior using quantitative Mueller matrix parameters. In the processes of rigor mortis and proteolysis of muscle samples, multispectral frequency distribution histograms (FDHs) of the Mueller matrix elements can reveal rich qualitative structural information. In addition, we analyze the temporal variations of the sample using the multispectral Mueller matrix transformation (MMT) parameters. The experimental results indicate that the different stages of rigor mortis and proteolysis for bovine skeletal muscle samples can be judged by these MMT parameters. The results presented in this work show that combining with the multispectral technique, the FDHs and MMT parameters can characterize the microstructural variation features of skeletal muscle tissues. The techniques have the potential to be used as tools for quantitative assessment of meat qualities in food industry.
Hyperspectral-stimulated Raman scattering imaging of cholesteryl ester accumulation: new avenue to diagnosis of human prostate cancer
Most prostate cancers (PCa) are slowly growing, and only the aggressive ones require early diagnosis and effective treatment. The current standard for PCa diagnosis remains histopathology. Nonetheless, for the differentiation between Gleason score 6 (low-risk PCa), which can be left without treatment, and Gleason score 7 (high-risk PCa), which requires active treatment, the inter-observer discordance can be up to 40%. Our previous study reveals that cholesteryl ester (CE) accumulation induced by PI3K/AKT activation underlies human PCa aggressiveness. However, Raman spectromicroscopy used in this study could only provide compositional information of certain lipid droplets (LDs) selected by the observer, which overlooked cell-to-cell variation and hindered translation to accurate automated diagnosis. Here, we demonstrated quantitative mapping of CE level in human prostate tissues using hyperspectral stimulated Raman scattering (SRS) microscopy that renders compositional information for every pixel in the image. Specifically, hundreds of SRS images at Raman shift between 1620-1800 cm-1 were taken, and multivariate curve resolution algorism was used to retrieve concentration images of acyl C=C bond, sterol C=C bond, and ester C=O bond. Given that the ratio between images of sterol C=C and ester C=O (sterol C=C/C=O) is nonlinearly proportional to CE percentage out of total lipid, we were able to quantitatively map CE level. Our data showed that CE level was significantly greater in high Gleason grade compared to low Gleason grade, and could be a factor that significantly contributed to cancer recurrence. Our study provides an opportunity towards more accurate PCa diagnosis and prediction of aggressiveness.
Poster Session
Hemoglobin concentration determination based on near infrared spatially resolved transmission spectra
Linna Zhang, Gang Li, Ling Lin
Spatially resolved diffuse reflectance spectroscopy method has been proved to be more effective than single point spectroscopy method in the experiment to predict the concentration of the Intralipid diluted solutions. However, Intralipid diluted solution is simple, cannot be the representative of turbid liquids. Blood is a natural and meaningful turbid liquid, more complicate. Hemoglobin is the major constituent of the whole blood. And hemoglobin concentration is commonly used in clinical medicine to diagnose many diseases. In this paper, near infrared spatially resolved transmission spectra (NIRSRTS) and Partial Least Square Regression (PLSR) were used to predict the hemoglobin concentration of human blood. The results showed the prediction ability for hemoglobin concentration of the proposed method is better than single point transmission spectroscopy method. This paper demonstrated the feasibility of the spatially resolved diffuse reflectance spectroscopy method for practical liquid composition analysis. This research provided a new thinking of practical turbid liquid composition analysis.
Research of transmissive near infrared spectroscopy for non-invasive blood glucose measurement
Near infrared (NIR) has prospectively applied in non-invasive blood glucose measurement due to glucose absorption among the 1.0-2.5m spectral bands. However, this significant technology is hard to be developed because of other blood components and low signal-to-noise ratio (SNR). In this work, we presented a non-invasive glucose measurement system using Fourier transform spectrometer which will work in fingertips or other human body tissues. A refrigerated InGaAs detector with high quantum efficiency performing well in the range of 1.0-1.7μm wavelength is used to acquire transmissive radiation. Preliminary experiment investigations were set up to test glucose levels of aqueous solutions with different concentrations. The analytical modeling of the interferogram data is based on arithmetic Fourier transform and supported by the curvilineal characterization. Experimental results show the variation of light intensity among different glucose concentrations and emphasize the obvious absorption of glucose in NIR wave-range. This study confirms the suitability that NIR can be developed in non-invasive glucose measurement.
Amplitude enhancement by a gold dimer
Xin Hong, Jingxin Wang, Zheng Jin
The unique optical properties such as brightness, non-bleaching, good bio-compatibility make gold particles ideal label candidates for molecular probes. Due to the strongly enhanced field, aggregation of gold nanoparticles finds themselves plenty of applications in bio-imaging. But limited by its small cross-section associated with nanometer sized particle, it is a big challenge to employ it in a single molecular detection. The field enhancement results from the effect of plasmonic coupling between two closely attached gold nanoparticle under the right excitation condition. With the aim to apply the gold dimer probe to find the molecules in our recently established optical detection method, we compared of the amplitude enhancement by the dimer relative to a single particle. The amplitude distribution under a highly focused illumination objective was calculated, whose results suggest that at the optimized excitation condition, the local field can be enhanced ~190 fold. In consequence, experimental detection was carried out. Gold dimers were linked together by the hybridization of two single chain DNAs. Dimer and single particle probes were mixed together in one detection. Overwhelming contrast between these two kinds of probes were clearly exhibited in the experimental detection image. This method can provide a way to a high specific detection in early diagnosis.
Wavelength selection based on two-dimensional correlation spectroscopy: application to noninvasive hemoglobin measurement by dynamic spectrum
Shengzhao Zhang, Linna Zhang, Zhe Li, et al.
Dynamic spectrum (DS) method is one of the noninvasive approaches to measure the concentration of components in human blood based on the application of photoplethysmogram (PPG). One of the targets of the DS method is to predict the hemoglobin concentration in human blood noninvasively. In previous works, the usually used wavelength in the spectrum is 600-1100 nm which is regarded as the analysis “window” in human tissues. Optimum wavelengths for measurements of hemoglobin concentration have not been investigated yet. In order to improve the precision and reliability of hemoglobin measurements, a method for wavelength selection based on two-dimension (2D) correlation spectroscopy has been studied in this paper. By analyzing the 2D correlation spectroscopy which is generated by the DS data from subject with different blood hemoglobin concentrations, the wavelength bands which are sensible to hemoglobin concentrations in DS can be found. We developed calibration models between the DS data and hemoglobin concentration based on data from 57 subjects. The correlation coefficient is 0.68 in the test set of the model using the whole wavelength band (600-1100nm), while in the test set of the model using the selected wavelength band (850- 950nm) the correlation coefficient is 0.87. Results show the feasibility of wavelength selection utilizing 2Dcorrelation spectroscopy.
Adaptive photoacoustic imaging quality optimization with EMD and reconstruction
Chengwen Guo, Yao Ding, Jie Yuan, et al.
Biomedical photoacoustic (PA) signal is characterized with extremely low signal to noise ratio which will yield significant artifacts in photoacoustic tomography (PAT) images. Since PA signals acquired by ultrasound transducers are non-linear and non-stationary, traditional data analysis methods such as Fourier and wavelet method cannot give useful information for further research. In this paper, we introduce an adaptive method to improve the quality of PA imaging based on empirical mode decomposition (EMD) and reconstruction. Data acquired by ultrasound transducers are adaptively decomposed into several intrinsic mode functions (IMFs) after a sifting pre-process. Since noise is randomly distributed in different IMFs, depressing IMFs with more noise while enhancing IMFs with less noise can effectively enhance the quality of reconstructed PAT images. However, searching optimal parameters by means of brute force searching algorithms will cost too much time, which prevent this method from practical use. To find parameters within reasonable time, heuristic algorithms, which are designed for finding good solutions more efficiently when traditional methods are too slow, are adopted in our method. Two of the heuristic algorithms, Simulated Annealing Algorithm, a probabilistic method to approximate the global optimal solution, and Artificial Bee Colony Algorithm, an optimization method inspired by the foraging behavior of bee swarm, are selected to search optimal parameters of IMFs in this paper. The effectiveness of our proposed method is proved both on simulated data and PA signals from real biomedical tissue, which might bear the potential for future clinical PA imaging de-noising.
Influence of incident light offset on diffuse reflectance measurement for curved object: a Monte Carlo-based study
Chizhu Ding
Diffuse reflectance spectroscopy in the near-infrared (NIR) spectral ranges is a widely used technique for nondestructive inspection of biological tissues. The optical properties, such as absorption and scattering coefficients, can be inversely deduced from the measured quantities and then be used to speculate on some related chemical and physical properties of the tissue. Most studies consider biological tissues as homogeneous semi-infinite turbid media or infinitelywide planar layered turbid media. However, the biological tissues have various geometries, and nearly all of them have curved surfaces. The position and direction of the incident light relative to the tissue surface affect the diffuse reflectance. In this work, we study the influence of incident light offset on the measured diffuse reflectance signals based on the Monte Carlo (MC) simulation. The MC method are regarded as golden standard for light propagation in turbid media and can be used without the limitations of complex tissue geometries. A model for diffuse reflectance spectroscopy measurement using optic fiber probe is built. The incident light is assumed to be an infinitely narrow photon beam. The tissue under detection is assumed to be spherical described by its curvature radius. A series of Monte Carlo simulation are carried out with varying incident directions. Simulation results are analyzed and discussed to assess the influence on the measurements for tissues with different curvature radii. This study may aid in achieving more accurate and effective measurement without extensive experiments.
Pigmented skin lesion detection using random forest and wavelet-based texture
The incidence of cutaneous malignant melanoma, a disease of worldwide distribution and is the deadliest form of skin cancer, has been rapidly increasing over the last few decades. Because advanced cutaneous melanoma is still incurable, early detection is an important step toward a reduction in mortality. Dermoscopy photographs are commonly used in melanoma diagnosis and can capture detailed features of a lesion. A great variability exists in the visual appearance of pigmented skin lesions. Therefore, in order to minimize the diagnostic errors that result from the difficulty and subjectivity of visual interpretation, an automatic detection approach is required. The objectives of this paper were to propose a hybrid method using random forest and Gabor wavelet transformation to accurately differentiate which part belong to lesion area and the other is not in a dermoscopy photographs and analyze segmentation accuracy. A random forest classifier consisting of a set of decision trees was used for classification. Gabor wavelets transformation are the mathematical model of visual cortical cells of mammalian brain and an image can be decomposed into multiple scales and multiple orientations by using it. The Gabor function has been recognized as a very useful tool in texture analysis, due to its optimal localization properties in both spatial and frequency domain. Texture features based on Gabor wavelets transformation are found by the Gabor filtered image. Experiment results indicate the following: (1) the proposed algorithm based on random forest outperformed the-state-of-the-art in pigmented skin lesions detection (2) and the inclusion of Gabor wavelet transformation based texture features improved segmentation accuracy significantly.
Wavelength-swept fiber laser based on acousto-optic tuning method
Ming-hui Chen, Yun-ping Fan, Hao Zhang, et al.
In this study, we have demonstrated a wavelength-swept fiber laser based on an acousto-optic tunable filter(AOTF) as a selective element and a semiconductor optical amplifier(SOA) as a gain medium in an internal fiber ring cavity. The light deriving from one port of the SOA goes through an optical isolator, the AOTF, a fiber coupler and a polarized controller successively, then it goes back to the other port of the SOA to form a ring cavity. The laser output is from another port of the fiber coupler. The laser made by this method is mainly used for swept-source optical coherence tomography(SS-OCT). The application of the SOA provides a sufficiently broad range and can ensure an increased axial resolution of SS-OCT. AOTF offers a wide tuning range, high switching speed and stable operation against vibration for the non-mechanical structure. The proposed wavelength-swept fiber laser ensures a high axial resolution of tomographic images and has a stable laser output. We have discussed the influence of the SOA injection current to the tuning range of the laser. In the SOA injection current of 280 mA, a continuous wavelength tuning range from 1295 to 1370 nm centered at a wavelength of 1330nm is obtained at the sweep rate of 1.06 kHz, and the power of the swept source was 1.14 mW. In addition, for quantitative characterization of the wavelength-swept performance with a AOTF, we have theoretically and experimentally analyzed the influence of the following controllable parameters: injection current, output power and sweeping frequency.
Identification of tumor cells infiltrating into connective tissue in esophageal cancer by multiphoton microscopy
Jian Xu, Liwei Jiang, Deyong Kang, et al.
Esophageal cancer is one of the most common malignancies of the gastrointestinal cancers and carries poorer prognosis than other gastrointestinal cancers. In general practice, the depth of tumor infiltration in esophageal wall is crucial to establishing appropriate treatment plan which is established by detecting the tumor infiltration depth. Connective tissue is one of the main structures that form the esophageal wall. So, identification of tumor cells infiltrating into connective tissue is helping for detecting the tumor infiltration depth. Our aim is to evaluate whether multiphoton microscopy (MPM) can be used to detect tumor cells infiltrating into connective tissue in the esophageal cancer. MPM is well-suited for real-time detecting morphologic and cellular changes in fresh tissues since many endogenous fluorophores of fresh tissues are excited through two-photon excited fluorescence (TPEF) and second harmonic generation (SHG). In this work, microstructure of tumor cells and connective tissue are first studied. Then, morphological changes of collagen fibers after the infiltration of tumor cells are shown. These results show that MPM has the ability to detect tumor cells infiltrating into connective tissue in the esophageal cancer. In the future, MPM may be a promising imaging technique for detecting tumor cells in esophageal cancer.
Identification of calcifications in intracranial neoplasms using two photon excitation fluorescence microscopy
Peihua Lin, Xingfu Wang, Zanyi Wu, et al.
Calcifications within brain tumors may be an indicator of a relatively long survival because a long time is required for the formation of calcium deposits, and may present a novel biomarker associated with response and improved outcome of therapy. In this paper, we describe the use of two-photon excitation fluorescent (TPEF) microscopy combined second harmonic generation (SHG) microscopy for high-resolution imaging that can be applied in identification of intratumoral calcifications. Our results demonstrate that the calcification has stronger TPEF signal than the area around it and the emission spectra shows the difference between the two areas clearly. The TPEF image of calcified region corresponds well with the corresponding H&E stained image. In this work, we present that the label-free imaging technique is able to distinguish the calcified mass lesions in intracranial neoplasms reliably.
Identification of the boundary between normal brain tissue and ischemia region using two-photon excitation fluorescence microscopy
Huiping Du, Shu Wang, Xingfu Wang, et al.
Ischemic stroke is one of the common neurological diseases, and it is becoming the leading causes of death and permanent disability around the world. Early and accurate identification of the potentially salvageable boundary region of ischemia brain tissues may enable selection of the most appropriate candidates for early stroke therapies. In this work, TPEF microscopy was used to image the microstructures of normal brain tissues, ischemia regions and the boundary region between normal and ischemia brain tissues. The ischemia brain tissues from Sprague-Dawley (SD) rats were subjected to 6 hours of middle cerebral artery occlusion (MCAO). Our study demonstrates that TPEF microscopy has the ability to not only reveal the morphological changes of the neurons but also identify the boundary between normal brain tissue and ischemia region, which correspond well to the hematoxylin and eosin (H and E) stained images. With the development of miniaturized TPEF microscope imaging devices, TPEF microscopy can be developed into an effectively diagnostic and monitoring tool for cerebral ischemia.
Quantification of collagen distributions in rat hyaline and fibro cartilages based on second harmonic generation imaging
Xiaoqin Zhu, Chenxi Liao, Zhenyu Wang, et al.
Hyaline cartilage is a semitransparent tissue composed of proteoglycan and thicker type II collagen fibers, while fibro cartilage large bundles of type I collagen besides other territorial matrix and chondrocytes. It is reported that the meniscus (fibro cartilage) has a greater capacity to regenerate and close a wound compared to articular cartilage (hyaline cartilage). And fibro cartilage often replaces the type II collagen-rich hyaline following trauma, leading to scar tissue that is composed of rigid type I collagen. The visualization and quantification of the collagen fibrillar meshwork is important for understanding the role of fibril reorganization during the healing process and how different types of cartilage contribute to wound closure. In this study, second harmonic generation (SHG) microscope was applied to image the articular and meniscus cartilage, and textural analysis were developed to quantify the collagen distribution. High-resolution images were achieved based on the SHG signal from collagen within fresh specimens, and detailed observations of tissue morphology and microstructural distribution were obtained without shrinkage or distortion. Textural analysis of SHG images was performed to confirm that collagen in fibrocartilage showed significantly coarser compared to collagen in hyaline cartilage (p < 0.01). Our results show that each type of cartilage has different structural features, which may significantly contribute to pathology when damaged. Our findings demonstrate that SHG microscopy holds potential as a clinically relevant diagnostic tool for imaging degenerative tissues or assessing wound repair following cartilage injury.
Label-free imaging of rat spinal cords based on multiphoton microscopy
Chenxi Liao, Zhenyu Wang, Linquan Zhou, et al.
As an integral part of the central nervous system, the spinal cord is a communication cable between the body and the brain. It mainly contains neurons, glial cells, nerve fibers and fiber tracts. The recent development of the optical imaging technique allows high-resolution imaging of biological tissues with the great potential for non-invasively looking inside the body. In this work, we evaluate the imaging capacity of multiphoton microscopy (MPM) based on second harmonic generation (SHG) and two-photon excited fluorescence (TPEF) for the cells and extracellular matrix in the spinal cord at molecular level. Rat spinal cord tissues were sectioned and imaged by MPM to demonstrate that MPM is able to show the microstructure including white matter, gray matter, ventral horns, dorsal horns, and axons based on the distinct intrinsic sources in each region of spinal cord. In the high-resolution and high-contrast MPM images, the cell profile can be clearly identified as dark shadows caused by nuclei and encircled by cytoplasm. The nerve fibers in white matter region emitted both SHG and TPEF signals. The multiphoton microscopic imaging technique proves to be a fast and effective tool for label-free imaging spinal cord tissues, based on endogenous signals in biological tissue. It has the potential to extend this optical technique to clinical study, where the rapid and damage-free imaging is needed.
A method based on coffee-ring deposition confocal Raman spectroscopy of analysis of melamine in milk
Zong Tan, Da Chen
In this work, an economical and high-efficiency method for detection of melamine in milk was developed. The enrichment effect of coffee-ring was combined with the micro-region analysis of confocal Raman spectroscopy, in addition, assisted with chemometric algorithmthe. Consequently, a desired result was obtained that the LOD of melamine in this method was 1 ppm, which was excellent because the sensitivity of conventional Raman detection was generally low. Furthermore, the whole process were processed in an easily available condition with almost no chemical reagents consumption, and the chosen substrates for the formation of coffee-ring were reusable. Thus, the method is environmental friendly and has a great potential application in food safety inspection.
Monitoring the elasticity changes of HeLa cells during mitosis by atomic force microscopy
Ningcheng Jiang, Yuhua Wang, Jinshu Zeng, et al.
Cell mitosis plays a crucial role in cell life activity, which is one of the important phases in cell division cycle. During the mitosis, the cytoskeleton micro-structure of the cell changed and the biomechanical properties of the cell may vary depending upon different mitosis stages. In this study, the elasticity property of HeLa cells during mitosis was monitored by atomic force microscopy. Also, the actin filaments in different mitosis stages of the cells were observed by confocal imaging. Our results show that the cell in anaphase is stiffer than that in metaphase and telophase. Furthermore, lots of actin filaments gathered in cells’ center area in anaphase, which contributes to the rigidity of the cell in this phase. Our findings demonstrate that the nano-biomechanics of living cells could provide a new index for characterizing cell physiological states.
Determination of acceptor-to-donor cross section ratio for two-photon excitation in living cells
The cross section is a significant parameter for fluorescence protein and determination of acceptor-to-donor cross section ratio for two-photon excitation in living cells is the vital issue for two-photon excitation FRET quantification. In this study, Hela cells were labeled with FPs that acceptor-to-donor concentration ratio is 1 to 1 and acceptor-to-donor cross section ratio ranged from 700nm to 960nm was obtained by emission spectral unmixing with independent excitation crosstalk correction. The results show that acceptor-to-donor cross section ratio declines with the excitation wavelength from 700nm to 790nm and then increases inversely from 790nm to 960nm. This method can quickly determine the cross section without any additional references, which could provide a powerful and convenient tool for measuring acceptor-to-donor cross section ratio by two-photon excitation in living cells.
Detection of mast cell secretion by using surface enhanced Raman scattering
Juan Li, Ren Li, Liqin Zheng, et al.
Acupuncture can cause a remarkable increase in degranulation of the mast cells, which has attracted the interest of researchers since the 1980s. Surface-enhanced Raman scattering (SERS) could obtain biochemical information with high sensitivity and specificity. In this study, SERS was used to detect the degree of degranulation of mast cells according to different incubate time. Mast cells was incubated with culture medium for 0 h, 12 h and 24 h, then centrifuge the culture medium, decant the supernatant, and discard the mast cell. SERS was performed to obtain the biochemical fingerprinting signatures of the centrifuged medium. The spectra data are then analyzed by spectral peaks attribution and the principal component analysis (PCA). The measured Raman spectra of the two groups were separated well by PCA. It indicated that mast cells had secreted some substances into cultured medium though degranulation did not happen.
Polymer dots with broadband optical absorption (500 nm - 700 nm) and high-efficiency photoacoustic conversion for in vivo multispectral photoacoustic imaging
Multi-spectral photoacoustic (MSPA) imaging can extract quantitative information of interesting component from morphological photoacoustic image of targets. A probe with an optimal optical absorption can be detected by MSPA imaging with higher sensitivity and specificity. Here we developed a Nano polymers dots (P-dots) for MSPA imaging that has a high photoacoustic conversion efficiency in a broad optical absorbing band (500 nm - 700 nm). In vivo MSPA imaging experiment was successfully performed in mouse model. The relative concentration map of P-dots was exactly obtained from the background of tissues, which demonstrated the potential use of P-dots in the bio-imaging field based on MSPA imaging.
Effect of 17[beta]-estradiol on the elasticity of MCF-7 cells by atomic force microscopy
Estrogen plays an important role in the development and progression of breast cancer, and it promotes proliferation, invasion and metastasis of breast cancer cells. In this paper, we investigated the effect of estrogen on the elasticity of breast cancer cells. 17β-estradiol, one of the most active estrogens in the human body was applied to MCF-7 living cells and the elasticity of breast cancer cells was measured by atomic force microscopy. The force spectroscopy was performed on the center of the cell and the Hertz model was used to calculate the elasticity modulus. Furthermore, the confocal fluorescence imaging was taken to observe the effect of 17β-estradiol on the actin distribution in the cells. The results show that the elasticity of the cells decreases rapidly after the addition of 17β-estradiol, which indicates that the cells appear softer for 17β-estradiol’s treatment. From the confocal imaging, it can be observed that the actin filament rearranged for 17β-estradiol’s treatment, which may lead to the alteration of the cell elasticity. Our findings may deepen our understanding on the rapid effect of 17β-estradiol to MCF-7 cells.
Numerical simulation and analysis of accurate blood oxygenation measurement by using optical resolution photoacoustic microscopy
Tianhao Yu, Qian Li, Lin Li, et al.
Accuracy of photoacoustic signal is the crux on measurement of oxygen saturation in functional photoacoustic imaging, which is influenced by factors such as defocus of laser beam, curve shape of large vessels and nonlinear saturation effect of optical absorption in biological tissues. We apply Monte Carlo model to simulate energy deposition in tissues and obtain photoacoustic signals reaching a simulated focused surface detector to investigate corresponding influence of these factors. We also apply compensation on photoacoustic imaging of in vivo cat cerebral cortex blood vessels, in which signals from different lateral positions of vessels are corrected based on simulation results. And this process on photoacoustic images can improve the smoothness and accuracy of oxygen saturation results.
In vivo noninvasive measurement of preprandial and postprandial blood glucose using optical coherence tomography
Ying Zhang, Xiyang Zhang, Zhifang Li, et al.
Blood glucose concentration measurement is essential for the diagnosis and treatment of diabetes. However, conventional glucose measurement methods are invasive and not suitable for real-time monitoring. This study demonstrated a noninvasive blood glucose measurement method using optical coherence tomography to image human lip in vivo. Optical coherence tomography (OCT) is a noninvasive and depth-resolved technique capable of acquiring tissue structure images in real time. Human lip has very thin skin and is full of blood vessels, which is appropriate for noninvasive glucose measurement. To verify the feasibility of OCT for glucose concentration monitoring, two groups of OCT imaging data were obtained from human lips of normal people. In one group, OCT images of lip were acquired from people on an empty stomach. In the other group, the same sites of lip were observed by OCT 2 hours after breakfast. Evident differences were found from two groups of OCT images that correspond to preprandial glucose and 2- hour postprandial glucose, respectively. The relationship between OCT image and blood glucose concentration was investigated. The result indicates that OCT possesses considerable prospects in terms of noninvasive blood glucose measurement.
Measuring blood oxygenation of pulsatile arteries using photoacoustic microscopy
Qian Li, Tianhao Yu, Lin Li, et al.
Heart pumps blood through the blood vessels to provide body with oxygen and nutrients. As the result, the blood flow, volume and oxygenation in arteries has a pulsatile nature. Measuring these pulsatile parameters enables more precise monitoring of oxygen metabolic rate and is thus valuable for researches and clinical applications. Photoacoustic microscopy (PAM) is a proven label-free method for in vivo measuring blood oxygenation at single blood vessel level. However, studies using PAM to observe the pulsatile nature of blood oxygenation in arteries were not reported. In this paper, we use optical-resolution PAM (OR-PAM) technology to study the blood oxygenation dynamics of pulsatile arteries. First, the ability of our OR-PAM system to accurately reflect the change of optical absorption in imaged objects is demonstrated in a phantom study. Then the system is used to image exposed cortical blood vessels of cat. The pulsatile nature of blood volume and oxygenation in arteries is clearly reflected in photoacoustic (PA) signals, whereas it’s not observable in veins. By using a multi-wavelength laser, the dynamics of the blood oxygenation of pulsatile arteries in cardiac cycles can be measured, based on the spectroscopic method.
Self-assembled dye-doped polymer microspheres as whispering gallery mode lasers
Xiaogang Chen, Hongyi Sun, Hongqin Yang, et al.
Microlasers based on high-Q whispering-gallery-mode (WGM) resonances are promising low-threshold laser sources for bio-sensing and imaging applications. In this talk, we demonstrate a cost effective approach to obtain size-controllable polymer microspheres, which can be served as good WGM microcavities. By injecting SU-8 solution into low-refractiveindex UV polymer, self-assembled spherical droplet with smooth surface can be created inside the elastic medium and then solidified by UV exposure. The size of the microspheres can be tuned from several to hundreds of microns. WGM Lasing has been achieved by optically pumping the dye-doped microspheres with ns lasers. Experimental results show that the microsphere lasers have high quality factors and low lasing thresholds. The self-assembled dye-doped polymer microspheres would provide an excellent platform for the micro-laser sources in on-chip biosensing and imaging systems.
A method to improve the measurement stability of scattering coefficients in lip with optical coherence tomography
Xiyang Zhang, Ying Zhang, Zhifang Li, et al.
Optical coherence tomography (OCT) is a non-invasive imaging technique that provides real-time two- and three-dimensional images of scattering tissues with micrometer resolution. Scattering coefficient is estimated based on the slope of OCT dependent of depth. However, stability of the OCT signal slope is dependent on tissue heterogeneity. Our work is to provide a method of combining post-processing Fourier filtration with automatic identification of fitting range to reduce the effect of tissue heterogeneity on scattering coefficient, and used for noninvasive monitoring of glucose concentration in lip. Axial and lateral resolutions of OCT are 20 μm and 15 μm respectively. OCT images were obtained from lip before breakfast and after breakfast respectively. Results demonstrate that the method improve the stability and the precision of measurement of scattering coefficient of lip. There are obviously different in scattering coefficient of tissue between preprandial glucose and 2-hours postprandial glucose. These pilot studies show that OCT scattering coefficient extraction of lip may be considered as a significant diagnostic marker. OCT has potential to monitor glucose-induced changes in lip in vivo.
The optimum measurement precision evaluation for blood components using near-infrared spectra on 1000-2500 nm
Ziyang Zhang, Di Sun, Tongshuai Han, et al.
In the non-invasive blood components measurement using near infrared spectroscopy, the useful signals caused by the concentration variation in the interested components, such as glucose, hemoglobin, albumin etc., are relative weak. Then the signals may be greatly disturbed by a lot of noises in various ways. We improved the signals by using the optimum path-length for the used wavelength to get a maximum variation of transmitted light intensity when the concentration of a component varies. And after the path-length optimization for every wavelength in 1000-2500 nm, we present the detection limits for the components, including glucose, hemoglobin and albumin, when measuring them in a tissue phantom. The evaluated detection limits could be the best reachable precision level since it assumed the measurement uses a high signal-to-noise ratio (SNR) signal and the optimum path-length. From the results, available wavelengths in 1000-2500 nm for the three component measurements can be screened by comparing their detection limit values with their measurement limit requirements. For other blood components measurement, the evaluation their detection limits could also be designed using the method proposed in this paper. Moreover, we use an equation to estimate the absorbance at the optimum path-length for every wavelength in 1000-2500 nm caused by the three components. It could be an easy way to realize the evaluation because adjusting the sample cell’s size to the precise path-length value for every wavelength is not necessary. This equation could also be referred to other blood components measurement using the optimum path-length for every used wavelength.
Quantitative optical biomarkers of lung cancer based intrinsic two-photon excited fluorescence signal
Jingwen Li, Zhenlin Zhan, Hongxin Lin, et al.
Alterations in the elastic fibers have been implicated in lung cancer. However, the label-free, microscopic imaging of elastic fibers in situ remains a major challenge. Here, we present the use of intrinsic two-photon excited fluorescence (TPEF) signal as a novel means for quantification of the elastic fibers in intact fresh human lung tissues. We obtained the TPEF images of elastic fibers from ex vivo the human lung tissues. We found that three features, including the elastic fibers area, the elastic fibers orientation, the elastic fibers structure, provide the quantitative identification of lung cancer and the direct visual cues for cancer versus non-cancer areas. These results suggest that the TPEF signal can be used as the label-free optical biomarkers for rapid clinical lung diagnosis and instant image-guided surgery.
Effects of large vessel on temperature distribution based on photothermal coupling interaction model
Zhifang Li, Xiyang Zhang, Zuoran Li, et al.
This paper is based on the finite element analysis method for studying effects of large blood vessel on temperature based on photothermal coupling interaction model, and it couples the physical field of optical transmission with the physical field of heat transfer in biological tissue by using COMSOL Multiphysics 4.4 software. The results demonstrate the cooling effect of large blood vessel, which can be potential application for the treatment of liver tumors.
Detection of the multiphoton signals in stained tissue using nonlinear optical microscopy
Yaping Zeng, Jian Xu, Deyong Kang, et al.
Multiphoton microscopy (MPM) based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) imaging, has become a powerful, important tool for tissue imaging at the molecular level. Recently, MPM is also used to image hematoxylin and eosin (H and E)-stained sections in cancer diagnostics. However, several studies have showed that the MPM images of tissue stained with H and E are significantly different from unstained tissue sections. Our aim was to detect of the multiphoton signals in stained tissue by using MPM. In this paper, MPM was used to image histological sections of esophageal invasive carcinoma tissues stained with H, E, H and E and fresh tissue. To detect of the multiphoton signals in stained tissue, the emission spectroscopic of tissue stained with H, E, H and E were obtained. For comparison, the fresh tissues were also investigated. Our results showed that the tissue stained with H, E, H and E could be detected by their TPEF signals. While the tissue stained with H and fresh tissue could be detected by their TPEF and SHG signals. In this work, we detect of the multiphoton signals in stained tissue. These findings will be useful for choosing suitable staining method so to improve the quality of MPM imaging in the future.
The novel drug delivery to vascular wall using laser driven thermal balloon: basic study ex vivo
To enhance drug delivery performance of popular drug eluting balloon against re-stenosis after angioplasty, we have an idea regarding to adjacent use of our unique laser driven thermal balloon of which characteristics could realize short term and uniform temperature elevation to modify drug delivery characteristics. We have already reported a delivery enhancement effect using this idea, however, detailed characteristics have not been studied yet. We studied balloon dilatation in terms of vascular circumferential tension on the heating drug delivery performance using porcine carotid artery wall ex vivo. The extracted carotid artery was used and circumferential tension of 0-30 mN/mm2 was added. Heating drug delivery was performed on this carotid artery with the heated solution of hydrophobic fluorescent Rhodamine B with 3 μg/ml in concentration at 37 and 70°C. We obtained a defined drug delivery quantity as well as delivery depth by a microscopic fluorescence measurement on a cross section of the drug delivered vessel wall. In the cases of 70°C, we found the drug penetration increase against 3°C case. We predict that the collagen thermal denaturation of the vessel wall may play important role to this penetration. In the case of 3°C, we found the drug concentration on the intimal surface with 7 mN/mm2 was increased as 10-30 times as other tension values. We found surface grooves in this case using an electron micrography. Therefore, we think that the drug delivery enhancement might be related to the groove formations of the vessel wall.
Optimization on source detector distance for the glucose sensing in a tissue phantom using near-infrared diffuse spectra
Chao Guo, Tongshuai Han, Ziyang Zhang, et al.
In the non-invasive blood glucose concentration (BGC) sensing, the measurement based on near infrared spectroscopy has been a promising technology since it had acquired dozens of satisfactory results in short-term glucose monitoring tests. However, it’s still necessary to improve the measurement precision because it has challenges of the reduced precision in a long-term test when a lot of variables in the test would exist. Considering the requirement of multivariable analysis, the signals of diffuse reflectance spectra should include enough absorption information from glucose. However, the sensitivity of diffuse light intensity to the absorption variation at different source detector separations (SDSs) could be different. We present an analysis method using Monte-Carlo (MC) simulation and the diffuse equation for reasonably selecting proper SDS to get a satisfactory glucose measurement precision when there are multivariable disturbances. In the case of measuring glucose in a tissue phantom using the waveband of 1000-1340 nm, we show the SDS optimization result by using this analysis method. The experiment was designed to measure the diffuse reflectance spectra at 0.1-3.0 mm with the step of 0.1 mm, and the phantom solutions with different glucose concentrations and hemoglobin concentrations are tested. The glucose prediction precision was evaluated using the root mean squared error of prediction (RMSEP) for the all SDSs of 0.1-3.0 mm, and the SDSs with the lower RMSEP were selected for use. Moreover, the selected SDSs in the experiment shows a similar conclusion from the MC simulation. This work could be referenced to the in vivo BGC measurement.
Assessment of spatial information for hyperspectral imaging of lesion
Xue Yang, Gang Li, Ling Lin
Multiple diseases such as breast tumor poses a great threat to women's health and life, while the traditional detection method is complex, costly and unsuitable for frequently self-examination, therefore, an inexpensive, convenient and efficient method for tumor self-inspection is needed urgently, and lesion localization is an important step. This paper proposes an self-examination method for positioning of a lesion.

The method adopts transillumination to acquire the hyperspectral images and to assess the spatial information of lesion. Firstly, multi-wavelength sources are modulated with frequency division, which is advantageous to separate images of different wavelength, meanwhile, the source serves as fill light to each other to improve the sensitivity in the low-lightlevel imaging. Secondly, the signal-to-noise ratio of transmitted images after demodulation are improved by frame accumulation technology. Next, gray distributions of transmitted images are analyzed. The gray-level differences is constituted by the actual transmitted images and fitting transmitted images of tissue without lesion, which is to rule out individual differences. Due to scattering effect, there will be transition zones between tissue and lesion, and the zone changes with wavelength change, which will help to identify the structure details of lesion. Finally, image segmentation is adopted to extract the lesion and the transition zones, and the spatial features of lesion are confirmed according to the transition zones and the differences of transmitted light intensity distributions. Experiment using flat-shaped tissue as an example shows that the proposed method can extract the space information of lesion.
Endoscopic optical coherence tomography using compressive sensing
Conventionally endoscopic OCT images are reconstructed by uniform sampling in both axial and transverse direction of the image. To shorten the scanning time and reduce the acquisition data, we propose to sparsely sample the axial scans and use compressive sensing (CS) to reconstruct the full endoscopic OCT image with randomly chosen axial scans that are much less than conventional Nyquist criteria requirement. And we study the ratio of sparse sampling numerically and experimentally that are required to reconstruct acceptable OCT images. We demonstrate that the OCT image acquisition time can be significantly reduced because of much less acquired data.
Temperature insensitive prediction of glucose concentration in turbid medium using multivariable calibration based on external parameter orthogonalization
Tongshuai Han, Ziyang Zhang, Cuiying Sun, et al.
The measurement accuracy of non-invasive blood glucose concentration (BGC) sensing with near-infrared spectroscopy is easily affected by the temperature variation in tissue because it would induce an unacceptable spectrum variation and the consequent prediction deviation. We use a multivariable correction method based on external parameter orthogonalization (EPO) to calibrate the spectral data recorded at different temperature values to reduce the spectral variation. The tested medium is a kind of tissue phantom, the Intralipid aqueous solution. The calibration uses a projection matrix to get the orthogonal spectral space to the variable of external parameter, i.e. temperature, and then the useful spectral information relative to glucose concentration has been reserved. Even more, training the projection matrix can be separated to building the calibration matrix for the prediction of glucose concentration as it only uses the representative samples’ data with temperature variation. The method presents a lower complexity than modeling a robust prediction matrix, which can be built from comprehensive spectral data involved the all variables both of BGC and temperature. In our test, the calibrated spectra with the same glucose concentration but different temperature values show a significantly improved repeatability. And then the glucose concentration prediction results show a lower root mean squared error of prediction (RMSEP) than that using the robust calibration model, which has considered the two variables. We also discuss the rationality of the representative samples chosen by EPO. This research may be referenced to the temperature calibration for in vivo BGC sensing.
Determination of the reference position in the near-infrared non-invasive blood glucose measurement in vivo
Guang Han, Jin Liu, Rong Liu, et al.
Position-based reference measurement method is taken as one of the most promising method in non-invasive measurement of blood glucose based on spectroscopic methodology. Selecting an appropriate source-detector separation as the reference position is important for deducting the influence of background change and reducing the loss of useful signals. Our group proposed a special source-detector separation named floating-reference position where the signal contains only background change, that is to say, the signal at this source-detector separation is uncorrelated with glucose concentration. The existence of floating-reference position has been verified in a three layer skin by Monte Carlo simulation and in the in vitro experiment. But it is difficult to verify the existence of floating-reference position on the human body because the interference is more complex during in vivo experiment. Aiming at this situation, this paper studies the determination of the best reference position on human body by collecting signals at several source-detector separations on the palm and measuring the true blood glucose levels during oral glucose tolerance test (OGTT) experiments of 3 volunteers. Partial least square (PLS) calibration model is established between the signals at every source-detector separation and its corresponding blood glucose levels. The results shows that the correlation coefficient (R) between 1.32 mm to 1.88 mm is lowest and they can be used as reference for background correction. The signal of this special position is important for improving the accuracy of near-infrared non-invasive blood glucose measurement.
Rotary-scanning optical resolution photoacoustic microscopy
Weizhi Qi, Lei Xi
Optical resolution photoacoustic microscopy (ORPAM) is currently one of the fastest evolving photoacoustic imaging modalities. It has a comparable spatial resolution to pure optical microscopic techniques such as epifluorescence microscopy, confocal microscopy, and two-photon microscopy, but also owns a deeper penetration depth. In this paper, we report a rotary-scanning (RS)-ORPAM that utilizes a galvanometer scanner integrated with objective to achieve rotary laser scanning. A 15 MHz cylindrically focused ultrasonic transducer is mounted onto a motorized rotation stage to follow optical scanning traces synchronously. To minimize the loss of signal to noise ratio, the acoustic focus is precisely adjusted to reach confocal with optical focus. Black tapes and carbon fibers are firstly imaged to evaluate the performance of the system, and then in vivo imaging of vasculature networks inside the ears and brains of mice is demonstrated using this system.
Evaluation on the detection limit of blood hemoglobin using photolepthysmography based on path-length optimization
Di Sun, Chao Guo, Ziyang Zhang, et al.
The blood hemoglobin concentration’s (BHC) measurement using Photoplethysmography (PPG), which gets blood absorption to near infrared light from the instantaneous pulse of transmitted light intensity, has not been applied to the clinical use due to the non-enough precision. The main challenge might be caused of the non-enough stable pulse signal when it’s very weak and it often varies in different human bodies or in the same body with different physiological states. We evaluated the detection limit of BHC using PPG as the measurement precision level, which can be considered as a best precision result because we got the relative stable subject’s pulse signals recorded by using a spectrometer with high signal-to-noise ratio (SNR) level, which is about 30000:1 in short term. Moreover, we optimized the used pathlength using the theory based on optimum pathlength to get a better sensitivity to the absorption variation in blood. The best detection limit was evaluated as about 1 g/L for BHC, and the best SNR of pulse for in vivo measurement was about 2000:1 at 1130 and 1250 nm. Meanwhile, we conclude that the SNR of pulse signal should be better than 400:1 when the required detection limit is set to 5 g/L. Our result would be a good reference to the BHC measurement to get a desired BHC measurement precision of real application.
Evaluation and recognition of skin images with aging by support vector machine
Liangjun Hu, Shulian Wu, Hui Li
Aging is a very important issue not only in dermatology, but also cosmetic science. Cutaneous aging involves both chronological and photoaging aging process. The evaluation and classification of aging is an important issue with the medical cosmetology workers nowadays. The purpose of this study is to assess chronological-age-related and photo-age-related of human skin. The texture features of skin surface skin, such as coarseness, contrast were analyzed by Fourier transform and Tamura. And the aim of it is to detect the object hidden in the skin texture in difference aging skin. Then, Support vector machine was applied to train the texture feature. The different age’s states were distinguished by the support vector machine (SVM) classifier. The results help us to further understand the mechanism of different aging skin from texture feature and help us to distinguish the different aging states.
Preliminary experiments on pharmacokinetic diffuse fluorescence tomography of CT-scanning mode
Yanqi Zhang, Xin Wang, Guoyan Yin, et al.
In vivo tomographic imaging of the fluorescence pharmacokinetic parameters in tissues can provide additional specific and quantitative physiological and pathological information to that of fluorescence concentration. This modality normally requires a highly-sensitive diffuse fluorescence tomography (DFT) working in dynamic way to finally extract the pharmacokinetic parameters from the measured pharmacokinetics-associated temporally-varying boundary intensity. This paper is devoted to preliminary experimental validation of our proposed direct reconstruction scheme of instantaneous sampling based pharmacokinetic-DFT: A highly-sensitive DFT system of CT-scanning mode working with parallel four photomultiplier-tube photon-counting channels is developed to generate an instantaneous sampling dataset; A direct reconstruction scheme then extracts images of the pharmacokinetic parameters using the adaptive-EKF strategy. We design a dynamic phantom that can simulate the agent metabolism in living tissue. The results of the dynamic phantom experiments verify the validity of the experiment system and reconstruction algorithms, and demonstrate that system provides good resolution, high sensitivity and quantitativeness at different pump speed.
Differentiation of highly metastatic nasopharyngeal carcinoma cells using multiphoton microscopy
Zhenlin Zhan, Zhenzhen Sun, Jingwen Li, et al.
The primary hypothesis tested in the study was that nasopharyngeal carcinoma (NPC) cells at different stage of invasion and metastasis can be differentiated using multiphoton microscopy (MPM). CNE1 and CNE2Z cells were cultured and used in this study. The activity of cell migration and invasion was measured using Transwell assays. At the same time, the morphologic features were quantified from the multiphoton images. The measurements of Transwell migration and invasion showed that the invasion and migration of CNE2Z cells were significantly enhanced when compared with that of CNE1 cells. Also, statistically significant differences in the morphologic features were found between two kinds of cancer cells. In conclusion, it is feasible to use MPM to differentiate cancer cells with different stage of invasion and metastasis.
Measuring the biomechanical properties of the actin in MCF-7 breast cancer cell with a combined system of AFM and SIM
Biomechanics of cell plays an important role in the behavior and development of diseases, which has a profound influence on the health, structural integrity, and function of cells. In this study, we proposed a method to assess the biomechanical properties in single breast cancer cell line MCF-7 by combining structured illumination microscopy (SIM) with atomic force microscopy (AFM). High resolution optical image of actin in MCF-7 cell and its elastography were obtained. The result shows that the quantitative resolution was improved by SIM, with 490 nm of conventional fluorescence image and 285 nm of reconstructed SIM image, which could give a precise location for AFM measurement. The elasticity of actin is about in the range of 10~1000 kPa. The proposed methods will be helpful in the understanding and clinical diagnosis of diseases at single cell level.
Childhood lymphoblastic leukemia adverse drug reactions: study of risk factors and therapy prognosis by optical methods
A. Zyubin, A. Lavrova, S. Babak, et al.
The treatment of acute lymphoblastic leukemia (ALL) can result in the side-effects such as kidney affection, hepatic failure and tissue hypoxia. We study dynamics of special biochemical marker of these pathologies - adenosine triphosphate, that is well-known substance of energy metabolism. We use methods of confocal microscopy for determining the cellular and mitochondrial concentration of adenosine triphosphate (ATP). Quantitative values of adenosine triphosphate were calculated for each patient and correlation with degree of side-effects had been done. The application of confocal microscopy for studying of side-effects and therapy of lymphoblastic leukemia is discussed.
Parameter estimation and analysis model selections in fluorescence correlation spectroscopy
Shiqing Dong, Jie Zhou, Xuemei Ding, et al.
Fluorescence correlation spectroscopy (FCS) is a powerful technique that could provide high temporal resolution and detection for the diffusions of biomolecules at extremely low concentrations. The accuracy of this approach primarily depends on experimental condition requirements and the data analysis model. In this study, we have set up a confocal-based FCS system. And then we used a Rhodamine6G solution to calibrate the system and get the related parameters. An experimental measurement was carried out on one-component solution to evaluate the relationship between a certain number of molecules and concentrations. The results showed FCS system we built was stable and valid. Finally, a two-component solution experiment was carried out to show the importance of analysis model selection. It is a promising method for single molecular diffusion study in living cells.
Acquisition of mouse optical structures in vivo with the aid of registered atlas
In fluorescence molecular tomography (FMT), the reconstruction results can greatly benefit from a priori information of accurate tissue optical-structures, which is difficult to be obtained in vivo with the traditional diffuse optical tomography (DOT) alone. One of the solutions is to apply a priori anatomical-structures obtained with anatomical imaging systems such as X-ray computed tomography (XCT) to constrain the reconstruction process of DOT. However, since the X-ray imaging mechanism limits the contrast between soft-tissues, it is difficult to segment the abdominal organs from XCT images. In order to overcome the challenges, the anatomical-structures of a target mouse are approximately obtained through registering a standard mouse anatomical atlas, i.e., the Digimouse, to its XCT volume with non-rigid image registration, and the optical-structures of the target mouse is approximately estimated through anatomical-structures guided time-resolve DOT strategy. Results of numerical simulations reveals that the an effective target atlas can be obtained through the registration method, and the results show that the absorption and reduced scattering coefficients of each organs can be recovered with reasonable accuracies.
Novel shadowless imaging for eyes-like diagnosis in vivo
Ning Xue, Kai Jiang, Qi Li, et al.
Eyes-like diagnosis was a traditional Chinese medicine method for many diseases, such as chronic gastritis, diabetes, hypertension etc. There was a close relationship between viscera and eyes-like. White-Eye was divided into fourteen sections, which corresponded to different viscera, so eyes-like was the reflection of status of viscera, in another words, it was an epitome of viscera health condition. In this paper, we developed a novel shadowless imaging technology and system for eyes-like diagnosis in vivo, which consisted of an optical shadowless imaging device for capturing and saving images of patients’ eyes-like, and a computer linked to the device for image processing. A character matching algorithm was developed to extract the character of white-eye in corresponding sections of eyes-like images taken by the optical shadowless imaging device, according to the character of eyes-like, whether there were viscera diseases could be learned. A series of assays were carried out, and the results verified the feasibility of eyes-like diagnosis technique.
Interstitial optical parameter quantification of turbid medium based on CW radiance measurements
CW radiance measurements examine the light intensity at a single source-detector location from different detection directions to recover absorption coefficient and reduced scattering coefficient of the turbid medium which is important in treatment planning of minimally invasive laser therapies. In this paper, P9 approximation for radiance is used as the forward model for fitting by considering the balance between computational time and the correctness of the forward model at low albedo and small source detector separation (SDS). By fitting P9 approximation for radiance to the angular radiance Monte Carlo (MC) simulations used as the angular radiance measurements, optical parameters are recovered over a wide range of reduced albedo between 0.69 and 0.99 at small SDS 2mm. The recovery errors of absorption coefficient and reduced scattering coefficient are less than 11.96% and 2.63%, respectively. The effects of the maximum angle used for fitting on optical parameter recovery have been further studied. The results show that the recovery errors of absorption coefficient and reduced scattering coefficient are less than 12% and 3% respectively when the maximum angle is greater than 70 degree.
Dorsal hand vein recognition based on Gabor multi-orientation fusion and multi-scale HOG features
Kinds of factors such as illumination and hand gestures would reduce the accuracy of dorsal hand vein recognition. Aiming at single hand vein image with low contrast and simple structure, an algorithm combining Gabor multi-orientation features fusion with Multi-scale Histogram of Oriented Gradient (MS-HOG) is proposed in this paper. With this method, more features will be extracted to improve the recognition accuracy. Firstly, diagrams of multi-scale and multi-orientation are acquired using Gabor transformation, then the Gabor features of the same scale and multi-orientation will be fused, and the features of the correspondent fusion diagrams will be extracted with a HOG operator of a certain scale. Finally the multi-scale cascaded histograms will be obtained for hand vein recognition. The experimental results show that our method not only improve the recognition accuracy but has good robustness in dorsal hand vein recognition.
Discrimination of liver cancer in cellular level based on backscatter micro-spectrum with PCA algorithm and BP neural network
Jing Yang, Cheng Wang, Gan Cai, et al.
The incidence and mortality rate of the primary liver cancer are very high and its postoperative metastasis and recurrence have become important factors to the prognosis of patients. Circulating tumor cells (CTC), as a new tumor marker, play important roles in the early diagnosis and individualized treatment. This paper presents an effective method to distinguish liver cancer based on the cellular scattering spectrum, which is a non-fluorescence technique based on the fiber confocal microscopic spectrometer. Combining the principal component analysis (PCA) with back propagation (BP) neural network were utilized to establish an automatic recognition model for backscatter spectrum of the liver cancer cells from blood cell. PCA was applied to reduce the dimension of the scattering spectral data which obtained by the fiber confocal microscopic spectrometer. After dimensionality reduction by PCA, a neural network pattern recognition model with 2 input layer nodes, 11 hidden layer nodes, 3 output nodes was established. We trained the network with 66 samples and also tested it. Results showed that the recognition rate of the three types of cells is more than 90%, the relative standard deviation is only 2.36%. The experimental results showed that the fiber confocal microscopic spectrometer combining with the algorithm of PCA and BP neural network can automatically identify the liver cancer cell from the blood cells. This will provide a better tool for investigating the metastasis of liver cancers in vivo, the biology metabolic characteristics of liver cancers and drug transportation. Additionally, it is obviously referential in practical application.
Compact hybrid real-time hyperspectral imaging system with high effective spatial, spectral, and temporal resolution
Filip Roth, Ahmad Abbadi, Ondrej Herman, et al.
Medical endoscopes for image-guided surgery commonly use standard color image sensors, discarding any more detailed spectral information. Medical spectroscopy devices with various spectral working ranges are specialized to specific medical procedures and in general are not usable for image-guided surgery due to limitations in spatial or temporal resolution.

In this paper, we present an initial demonstrator of hyperspectral endoscope, composed of two image sensors with complementing parameters. Using this hybrid approach, combining sensors with different spatial and spectral resolutions and spectral ranges, we obtain improved coverage of all the respective parameters. After digitally processing and merging the video streams, while maintaining the better features of both, we obtain an imaging system providing high effective spatial, spectral, and temporal resolution. The system is based on field programmable gate arrays. It provides real-time video output (60 Hz), which is usable for navigation during image-guided surgery. The flexible system architecture allows for an easy extension of the processing algorithms and enables minimal video signal latency. Physical dimensions and portability of the system are comparable to standard off-the-shelf medical endoscope cameras. The device can output both processed video and standard visible light video signals on one or more video outputs of the system. The resulting processed video signal obtained from the combined image sensor data greatly increases the amount of useful information available to the end user.
Interferometer immunosensor based on porous silicon for determining alpha-fetoprotein
Xiaoyi Lv, Jing Jiang, Guodong Lv, et al.
An increased level of alpha-fetoprotein ( AFP) in the blood may be a sign of liver cancer. Porous silicon based optical microcavities structure is prepared as a label-free immunosensor platform for detecting AFP. After the antigen-antibody reaction, it is monitored that the red shift of the reflection spectrum of the immunosensor increases
Label-free and high-sensitive detection for genetic point mutation based on hyperspectral interferometry
Rongxin Fu, Qi Li, Junqi Zhang, et al.
Label free point mutation detection is particularly momentous in the area of biomedical research and clinical diagnosis since gene mutations naturally occur and bring about highly fatal diseases. In this paper, a label free and high sensitive approach is proposed for point mutation detection based on hyperspectral interferometry. A hybridization strategy is designed to discriminate a single-base substitution with sequence-specific DNA ligase. Double-strand structures will take place only if added oligonucleotides are perfectly paired to the probe sequence. The proposed approach takes full use of the inherent conformation of double-strand DNA molecules on the substrate and a spectrum analysis method is established to point out the sub-nanoscale thickness variation, which benefits to high sensitive mutation detection. The limit of detection reach 4pg/mm2 according to the experimental result. A lung cancer gene point mutation was demonstrated, proving the high selectivity and multiplex analysis capability of the proposed biosensor.
Application study of transport intensity equation in quantitative phase reconstruction
Xiaojun Song, Wei Cheng, Chunjuan Wei, et al.
In order to improve detection speed and accuracy of biological cells, a quantitative non-interference optical phase recovery method is proposed in commercial microscope, taking the red blood cells as the classical phase objects. Three bright field micrographs were collected in the experiment. Utilizing the transport intensity equation (TIE), the quantitative phase distributions of red blood cell are gained and agree well with the previous optical phase models. Analysis shows that the resolution of introduced system reaches sub-micron. This method not only quickly gives quantitative phase distribution of cells, but also measures a large number of cells simultaneously. So it is potential in the use of real-time observing and quantitative analyzing of cells in vivo.
Photophysical property of the pyridyl and pyrimidinyloxy silicon (IV) phthalocyanines and their morphology of polymeric nanoparticles
Sujuan Pan, Zhe Chen, Shijun Wu, et al.
Phthalocyanines (Pcs) are extensively studied by many scientists because of their interesting optical, electrical properties, and good thermal stability. The unsubstituted Pcs can present solubility and aggregation behaviour problems for their limiting applications. In our study two pyridyl and pyrimidinyloxy silicon (IV) phthalocyanines were synthesized. Their photophysical properties were examined by UV-Vis, steady-state and time-resolved fluorescence spectroscopic methods. The positions of Q band were observed at 670 nm for two phthalocyanines. Compared with silicon phthalocyanine dichloride (SiPcCl2), the fluorescence intensities and lifetimes of pyridyl and pyrimidinyloxy silicon (IV) phthalocyanines increased. In order to improve biocompatibility and tumor-targeted delivery, the hydrophobic dendritic phthalocyanine were encapsulated by diblock amphiphilic copolymer poly (N’-benzyl oxygen carbonyl lysine)-poly (ethylene glycol)-poly (N’-benzyl oxygen carbonyl lysine) (PLL(Z)-PEG-PLL(Z)) to form the polymeric nanoparticles. The morphology of two nanoparticles were investigated by using atomic force microscope. The polymeric nanoparticles were spherical with the diameter at about 35 nm. The polymeric nanoparticle SiPc(OR2)2@PLL(Z)-PEG-PLL(Z) would be the promising third-generation photosensitizer (PS) for photodynamic therapy (PDT).
2D light scattering label-free cytometry using light-sheet illumination
Two-dimensional (2D) light scattering cytometry has been demonstrated as an effective label-free technology for cell analysis. Here we develop the light-sheet illumination in 2D light scattering static cytometry. In our cytometer, a cylindrical lens is used to form the light-sheet for better excitation of the static cells under an inverted microscope. The thickness of the light-sheet measured in fluorescent solution is about 13 μm. Two-dimensional light scattering patterns of standard microspheres and yeast cells are obtained by using a complementary metal oxide semiconductor (CMOS) detector via a low numerical aperture (NA 0.4) optical objective. The experimental patterns characterized with fringe structures agree well with Mie theory simulated ones. Our results suggest that the light-sheet illumination is an effective excitation method for 2D light scattering label-free cytometry.
The effect of metal ions on the photophysical and photochemical property of phenylthio bromo metal phthalocyanines
Sujuan Pan, Pingping Shen, Dongdong Ma, et al.
Phthalocyanines have attracted great attention because of their applications in material science including electro-optical devices, electrochromic display, and photodynamic therapy (PDT) of cancer. In addition, the Pcs exhibit great flexibility of chemical structure modification enabled by either peripheral substituents or metal ions co-ordination to central cavity of highly conjugated tetrapyrrolic macrocycles. However, because of the hydrophobic nature of the phthalocyanine ring, Pcs have strong tendency to aggregate in solution, which limited their applications. To overcome this problem, the introduction of dendritic wedge to peripheral positions of phthalocyanines can prevent the formation of aggregation to some extent. The preparation procedure involved the modification of the zinc (II) and magnesium (II) phthalocyanines with peripherally dendritic substitutions. The photophysical and photochemical properties of dendritic phthalocyanines were studied by UV/Vis and fluorescence spectroscopic methods. Compared with the magnesium (II) phthalocyanine, the intensity of Q band of zinc (II) phthalocyanine was increased but no obviously position changes was observed. Furthermore, the zinc (II) phthalocyanine exhibited relatively higher fluorescence intensity than the magnesium (II) phthalocyanine. The fluorescence quantum yield and lifetimes of magnesium (II) phthalocyanine was clearly longer than that of zinc (II) phthalocyanine. As the better photosensitizer, the zinc (II) phthalocyanine has higher singlet oxygen quantum yield owning superior performance. This results indicated that the singlet oxygen quantum yield would be effected by the nature of metal ions.
Monitoring the change of mitochondrial morphology and its metabolism of the breast cancer cells with the treatment of Hsp70 inhibitor during heat shock by fluorescence imaging
Heat shock (HS) is one of the best-studied exogenous cellular stresses, and all cellular compartments and metabolic processes are involved in HS response. The heat shock proteins (Hsps) expression enhanced during HS mainly localized in subcellular compartments, such as cytosol, endoplasmic reticulum and mitochandria. The major inducible heat shock protein 70 (Hsp70) modulate cellular homeostasis and promote cellular survival by blocking a caspase independent cell death through its association with apoptosis inducing factor. Mitochondria as the critical elements of HS response that participate in key metabolic reactions, and the changes in mitochonrial morphology may impact on mitochondrial metabolism. In this paper, the changes of mitorchondrial morphology in breast cancer cell have been monitored in real time after heat shock (43 ℃ ) by the fluorescence imaging, and the influence of Hsp70 inhibitor on mitochandrial structures have also been investigated. Then the information of mitochondrial metabolism which can be characterized by the level of the mitochondrial membrane potential has also been obtained wihout/with the treatment of Hsp70 inhibitor. Our data indicated that the mitochandrial morphology were related with the mitochandrial membrane potential, and the mitochandrial membrane potential was influenced significantly with the treatment of Hsp70 inhibitor during HS.
Development of wide-angle 2D light scattering static cytometry
Linyan Xie, Qiao Liu, Changshun Shao, et al.
We have recently developed a 2D light scattering static cytometer for cellular analysis in a label-free manner, which measures side scatter (SSC) light in the polar angular range from 79 to 101 degrees. Compared with conventional flow cytometry, our cytometric technique requires no fluorescent labeling of the cells, and static cytometry measurements can be performed without flow control. In this paper we present an improved label-free static cytometer that can obtain 2D light scattering patterns in a wider angular range. By illuminating the static microspheres on chip with a scanning optical fiber, wide-angle 2D light scattering patterns of single standard microspheres with a mean diameter of 3.87 μm are obtained. The 2D patterns of 3.87 μm microspheres contain both large-angle forward scatter (FSC) and SSC light in the polar angular range from 40 to 100 degrees, approximately. Experimental 2D patterns of 3.87 μm microspheres are in good agreement with Mie theory simulated ones. The wide-angle light scattering measurements may provide a better resolution for particle analysis as compared with the SSC measurements. Two dimensional light scattering patterns of HL-60 human acute leukemia cells are obtained by using our static cytometer. Compared with SSC 2D light scattering patterns, wide-angle 2D patterns contain richer information of the HL-60 cells. The obtaining of 2D light scattering patterns in a wide angular range could help to enhance the capabilities of our label-free static cytometry for cell analysis.
Diffuse reflectance spectroscopy study of in vitro tissue for nasopharyngeal carcinoma diagnosis
Diffuse reflectance spectroscopy is a non-contact, non-invasive, and low-cost optical technique that provides real-time feedback about the absorptive characteristics and the microstructure properties of biological tissue. This optical technique shows the potential for monitoring metabolic status associated with malignancy transformation. Nasopharyngeal carcinoma (NPC) is the third most frequently diagnosed cancer associated with virus and is the most common male malignancy with a characteristic regional and racial distribution worldwide. This paper investigates the current screening state of nasopharyngeal malignancies and also provides an overview on the applications of diffuse reflectance spectroscopy in the cancer detection. Furthermore, the latest research relevant to the diagnosis of NPC in vitro tissue using diffuse reflectance spectroscopy is introduced. The results of diffuse reflectance spectroscopy are summarized, showing a significant experimental and clinical value for further NPC detection in vivo in the future.
Photophysical properties of catechol axially substituted tetra-[alpha]-(pentyloxy) titanium (IV) phthalocyanine
Xinxin Yu, Ting Lin, Huafei Lv, et al.
Metal phthalocyanines (MPcs) have been found to be a promising photosensitizers for photodynamic therapy (PDT) of cancers and non-cancer diseases. Nevertheless, phthalocyanines are substantially limited in clinical applications owing to their poor solubility, aggregation and insufficient selectivity for cancer cells. Catechol is an important pharmaceutical intermediate, playing important in vivo biological activity in medicine. Using catechol (pyrocatechin) as axial ligands, utilizing of the pharmaceutical effect of catechins, could improve the bioavailability, and achieve synergistic therapeutic effect in PDT. To address these issues, a novel catechol axially substituted tetra-α-(pentyloxy) titanium(IV) (TiPc(OC5H11)4-Catechol) was synthesized. The structure of TiPc(OC5H11)4-Catechol was characterized by elemental analysis, IR, 1HNMR and MS methods. The photophysical properties of TiPc(OC5H11)4 and TiPc(OC5H11)4-Catechol have been studied by UV/Vis and steady-state fluorescence spectra. After being axially substituted with catechin groups, no obviously intensity and position of maximum wavelength in Q-band of TiPc(OC5H11)4 and TiPc(OC5H11)4-Catechol were observed. The fluorescence intensity of TiPc(OC5H11)4 was stronger than that of TiPc(OC5H11)4-Catechol, but the fluorescence lifetime of TiPc(OC5H11)4-Catechol was longer than that of TiPc(OC5H11)4. TiPc(OC5H11)4-Catechol may be considered as a promising photosensitizer for PDT.
Integrated acoustic-resolution and optical-resolution photoacoustic microscopy using a single multifunctional acoustic lens
Heng Guo, Lei Xi
With the rapid development of photoacoustic imaging, it has been widely used in various research fields such as biology, medicine and nanotechnology. Due to the huge difference among photoacoustic imaging systems, it is hard to integrate them in one platform. To solve this problem, we propose to develop a new universal photoacoustic imaging platform that integrates acoustic-resolution photoacoustic microscopy and optical-resolution photoacoustic microscopy through a multifunctional liquid lens. This lens takes advantage of an inherently low acoustic impedance and a tunable focal length that was characterized by the infusion volume of the liquid. In this paper, the liquid lens was used to realize confocal of laser illumination and acoustic detection for both acoustic-resolution and optical-resolution photoacoustic microscopy. The home-made polyvinylidene fluoride (PVDF) acoustic transducer had a center frequency of 10MHz and -6dB frequency spectrum from 4MHz to 15MHz which yielded to an axial resolution of 70 μm. The lateral resolutions of acoustic- and optical-resolution photoacoustic microscopy were evaluated to be 180 μm and 4.8 μm, respectively. The vasculature of rat ears was carried out to evaluate the performance of optical-resolution photoacoustic microscopy.
En-face sectional imaging using single-shot full-field optical coherence tomography (SS-FF-OCT) based on white light emitting diode (WLED)
Tulsi Anna, Chih-Ming Lai, Arthur Chiou, et al.
This work reports a Linnik type single shot full-field optical coherence tomography (SS-FF-OCT), which uses a fast generalized analytic signal based complex Riesz transform scheme to reconstruct wide area en-face OCT images. The OCT interferometer is illuminated using a single broad band white light emitting diode (WLED) (wavelength range 470- 850nm, central wavelength 650nm) and detection unit is a two-dimensional (2D) charge complementary metal oxide semiconductor (CMOS) camera. The measured axial and lateral resolution (using 4x, NA=0.1 microscope objective) of the present system is 0.9μm and 3.9μm, very close to theoretical values. The measured imaging area using CMOS camera is 2150 x 2150μm2. The feasibility of the system is demonstrated by imaging scattering samples such as onion bulk and plant leaf. The present SS-FF-OCT is compact, fast (Riesz transform based scheme), stable, cost-effective, and provides comparable axial resolution.
Whole-body and multispectral photoacoustic imaging of adult zebrafish
Na Huang, Lei Xi
Zebrafish is a top vertebrate model to study developmental biology and genetics, and it is becoming increasingly popular for studying human diseases due to its high genome similarity to that of humans and the optical transparency in embryonic stages. However, it becomes difficult for pure optical imaging techniques to volumetric visualize the internal organs and structures of wild-type zebrafish in juvenile and adult stages with excellent resolution and penetration depth. Even with the establishment of mutant lines which remain transparent over the life cycle, it is still a challenge for pure optical imaging modalities to image the whole body of adult zebrafish with micro-scale resolution. However, the method called photoacoustic imaging that combines all the advantages of the optical imaging and ultrasonic imaging provides a new way to image the whole body of the zebrafish. In this work, we developed a non-invasive photoacoustic imaging system with optimized near-infrared illumination and cylindrical scanning to image the zebrafish. The lateral and axial resolution yield to 80 μm and 600 μm, respectively. Multispectral strategy with wavelengths from 690 nm to 930 nm was employed to image various organs inside the zebrafish. From the reconstructed images, most major organs and structures inside the body can be precisely imaged. Quantitative and statistical analysis of absorption for organs under illumination with different wavelengths were carried out.
Revealing the cellular metabolism and microstructural changes in vivo in senescing Acer saccharum leaves using two-photon FLIM and full-field OCM
Seasonal as well as climate changes have immense effect on bud burst, leaf color and leaf abscission. Autumn phenology of leaves is clearly distinguishable in deciduous plant leaves where the leaf color changes from green to red (leaf senescence). In this work, two-photon fluorescence lifetime imaging microscopy (2P-FLIM) and full-field optical coherence microscopy (FF-OCM) were applied to study mitochondrial activity and microstructural changes, respectively, in the senescence of Acer saccharum (Sugar maple) leaves. Fluorescence lifetime of reduced nicotinamide adenine dinucleotide phosphate [NAD(P)H] was recorded using 2P-FLIM to quantify the cellular metabolic changes. Compared to the green leaves, the red leaves showed a 19% increase (P < 0.05) in the average fluorescence lifetime of NAD(P)H, and a 52% decrease (p < 0.005) in the free to protein-bound NAD(P)H ratio. This infers a significant change in mitochondrial metabolic regulation in red leaves in contrast to green leaves. Additionally, en-face sectional images at 0.8 μm axial resolutions of the green and the red color Acer saccharum leaves via FF-OCM using white light emitting diode (WLED) showed a well-defined microstructure of epicuticular waxy layer in green leaves as compared to red leaves where disintegrated microstructure was observed. Our approach can potentially be used to correlate mitochondrial activity with epicuticular microstructural changes in senescing leaves and other biological tissues.
Temporal evolution of liquid-assisted hard bio-tissue ablation with infrared pulsed lasers under a liquid environment
Faner Chen, Qiang Li, Mingxin Hua, et al.
Liquid-assisted hard biotissue ablation with the pulsed lasers takes advantages in precision and compatibility than mechanical tools in traditional surgery. The objective of this study was to monitor the dynamic process of the cavitation bubble evolution induced by Ho:YAG laser under water and identify the opening time of channel formation between the fiber tip to the target tissue surface. A free-running Ho:YAG laser was used in the experiment. The wavelength was 2.1 μm with a pulse duration of 350 us and pulse energy varied from 500 mJ to 2000 mJ. The high-speed camera (PCO. dimax, Germany, PCO) applied to monitor the whole ablation process was setting at a frame rate of 52000 frames/s. The results showed that the cavitation bubble induced by laser energy experienced an oscillation process including occurrence, expansion, contraction and subsequent collapse. A channel connected the fiber tip and target tissue surface was formed during the dynamic process which allowed the following pulse energy transmitted through the channel with a relative low absorption and directly interacted with the target tissue. The beginning time of channel formation, as well as the duration of channel opening, as functions of incident laser energy were also presented. A micro-explosion was observed near the tissue surface during the bubble collapse, which may contribute to produce a clean cut, reduce the thermal injury and improve the morphology of ablation crater.
A comparative study of metabolic state of stem cells during osteogenic and adipogenic differentiations via fluorescence lifetime imaging microscopy
Sandeep Chakraborty, Meng-Hsin Ou, Jean-Cheng Kuo, et al.
Cellular metabolic state can serve as a biomarker to indicate the differentiation potential of stem cells into other specialized cell lineages. In this study, two-photon fluorescence lifetime imaging microscopy (2P-FLIM) was applied to determine the fluorescence lifetime and the amounts of the auto-fluorescent metabolic co-factor reduced nicotinamide adenine dinucleotide (NADH) to elucidate the cellular metabolism of human mesenchymal stem cells (hMSCs) in osteogenic and adipogenic differentiation processes. 2P-FLIM provides the free to protein-bound NADH ratio which can serve as the indicator of cellular metabolic state. We measured NADH fluorescence lifetime at 0, 7, and 14 days after hMSCs were induced for either osteogenesis or adipogenesis. In both cases, the average fluorescence lifetime increased significantly at day 14 (P < 0.001), while the ratio of free to protein-bound NADH ratio decreased significantly in 7- days (P < 0.001) and 14-days (P < 0.001). Thus, our results indicated a higher metabolic rate in both osteogenic and adipogenic differentiation processes when compared with undifferentiated hMSCs. This approach may be further utilized to study proliferation efficiency and differentiation potential of stem cells into other specialized cell lineages.
Saliva surface-enhanced Raman spectroscopy for noninvasive optical detection of nasopharyngeal cancer
The early cancer detection is of great significance to increase the patient’s survival rate and reduce the risk of cancer development. Surface enhanced Raman spectroscopy (SERS) technique, a rapid, convenient, nondestructive optical detection method, can provide a characteristic “fingerprint” information of target substances, even achieving single molecule detection. Its ultra-high detection sensitivity has made it become one of the most potential biochemical detection methods. Saliva, a multi-constituent oral fluid, contains the bio-markers which is capable of reflecting the systemic health condition of human, showing promising potential as an effect medium for disease monitoring. Compared with the serum samples, the collection and processing of saliva is safer, more convenient and noninvasive. Thus, saliva test is becoming the hotspot issues of the noninvasive cancer research field. This review highlights and analyzes current application progress within the field of SERS saliva test in cancer detection. Meanwhile, the primary research results of SERS saliva for the noninvasive differentiation of nasopharyngeal cancer, normal and rhinitis obtained by our group are shown.
The effect of axial ligands on the quantum yield of singlet oxygen of new silicon phthalocyanine
Huafei Lv, Xuemei Zhang, Xinxin Yu, et al.
The singlet oxygen (1O2) production abilitity is an important factor to assess their potential as effective of photosensitizers. In this paper, the 1O2 production rate, production rate constant and quantum yield of silicon(IV) phthalocyanine axially bearing 1-3 generation dendritic substituents were evaluated by a high performance liquid chromatographic method. The results show that the 1O2 production rate and production rate constant of these compounds increase gradually with dendritic generations increase. And the 1O2 quantum yield of silicon(IV) phthalocyanine with first generation dendritic ligand was the highest. This may be due to the isolation effect of the dendritic ligands on the phthalocyanine core. The parameters of the observed 1O2 production properties will provide valuable data for these dendrimer phthalocyanines as promising photosensitizer in PDT application.
Photoinduced electron transfer between benzyloxy dendrimer phthalocyanine and benzoquinone
Tiantian Zhang, Dongdong Ma, Sujuan Pan, et al.
Photo-induced electron transfer (PET) is an important and fundamental process in natural photosynthesis. To mimic such interesting PET process, a suitable donor and acceptor couple were properly chosen. Dendrimer phthalocyanines and their derivatives have emerged as promising materials for artificial photosynthesis systems. In this paper, the electron transfer between the light harvest dendrimer phthalocyanine (donor) and the 1,4-benzoquinone (acceptor) was studied by UV/Vis and fluorescence spectroscopic methods. It was found that fluorescence of phthalocyanine was quenched by benzoquinone (BQ) via excited state electron transfer, from the phthalocyanine to the BQ upon excitation at 610 nm. The Stern–Volmer constant (KSV) of electron transfer was calculated. Our study suggests that this dendritic phthalocyanine is an effective new electron donor and transmission complex and could be used as a potential artificial photosynthesis system.
Study on nasopharyngeal cancer tissue using surface-enhanced Raman spectroscopy
Surface-enhanced Raman spectroscopy (SERS) can provide detailed molecular structure and composition information, and has demonstrated great potential in biomedical filed. This spectroscopy technology has become one of the most important optical techniques in the early diagnosis of cancer. Nasopharyngeal cancer (NPC) is a malignant neoplasm arising in the nasopharyngeal epithelial lining, which has relatively high incidence and death rate in Southeast Asia and southern China. This paper reviews the current progress of SERS in the field of cancer diagnostics, including gastric cancer, colorectal cancer, cervical cancer and nasopharyngeal cancer. In addition to above researches, we recently develop a novel NPC detection method based on tissue section using SERS, and obtain primary results. The proposed method has promising potential for the detection of nasopharyngeal carcinoma.
In vivo hyperspectral imaging and differentiation of skin cancer
Larisa A. Zherdeva, Ivan A. Bratchenko, Oleg O. Myakinin, et al.
Results of hyperspectral imaging analysis for in vivo visualization of skin neoplasms are presented. 16 melanomas, 19 basal cell carcinomas and 10 benign tumors with different stages of neoplasm growth were tested. The HSI system provide skin tissue images with 5 nm spectral resolution in the range of 450-750 nm with automatic stabilization of each frame compensating displacement of the scanning area due to spontaneous macro-movements of the patient. The integrated optical densities in 530-600 and 600-670 nm ranges are used for real-time hemoglobin and melanin distribution imaging in skin tissue. It was shown that the total accuracy of skin cancer identification exceeds 90% and 70% for differentiation of melanomas from BCC and begihn tumors. It was demonstrated the possibility for HSI classification of melanomas of different stages.
Skin cancer texture analysis of OCT images based on Haralick, fractal dimension, Markov random field features, and the complex directional field features
Dmitry S. Raupov, Oleg O. Myakinin, Ivan A. Bratchenko, et al.
In this paper, we propose a report about our examining of the validity of OCT in identifying changes using a skin cancer texture analysis compiled from Haralick texture features, fractal dimension, Markov random field method and the complex directional features from different tissues. Described features have been used to detect specific spatial characteristics, which can differentiate healthy tissue from diverse skin cancers in cross-section OCT images (B- and/or C-scans). In this work, we used an interval type-II fuzzy anisotropic diffusion algorithm for speckle noise reduction in OCT images. The Haralick texture features as contrast, correlation, energy, and homogeneity have been calculated in various directions. A box-counting method is performed to evaluate fractal dimension of skin probes. Markov random field have been used for the quality enhancing of the classifying. Additionally, we used the complex directional field calculated by the local gradient methodology to increase of the assessment quality of the diagnosis method. Our results demonstrate that these texture features may present helpful information to discriminate tumor from healthy tissue. The experimental data set contains 488 OCT-images with normal skin and tumors as Basal Cell Carcinoma (BCC), Malignant Melanoma (MM) and Nevus. All images were acquired from our laboratory SD-OCT setup based on broadband light source, delivering an output power of 20 mW at the central wavelength of 840 nm with a bandwidth of 25 nm. We obtained sensitivity about 97% and specificity about 73% for a task of discrimination between MM and Nevus.
Grid-based visual aid for enhanced microscopy screening in diagnostic cytopathology
Christos Riziotis, Evangelos Tsiambas
A grid acting as a spatial reference and calibration aid, fabricated on glass cover slips by laser micromachining and attached on the carrier microscope slide, is proposed as a visual aid for the improvement of microscopy diagnostic procedure in the screening of cytological slides. A set of borderline and also abnormal PAP test cases -according to Bethesda 2014 revised terminology- was analyzed by conventional and grid based screening procedures, and statistical analysis showed that the introduced grid-based microscopy led to an improved diagnosis by identifying an increased number of abnormal cells in a shorter period of time, especially concerning the number of pre- or neoplastic/cancerous cells.
Blood vessel damage correlated with irradiance for in vivo vascular targeted photodynamic therapy
Jinde Zhang, Zou Tan, Xiangyu Niu, et al.
Vascular targeted photodynamic therapy (V-PDT) has been widely utilized for the prevention or treatment of vascular-related diseases, including age-related macular degeneration, port-wine stains and prostate cancer. In order to quantitative assessment the blood vessel damage during V-PDT, nude mice were implanted with Titanium dorsal skin window chambers for in vivo V-PDT studies. For treatments, various irradiances including 50, 75, 100 and 200 mW/cm2 provided by a 532 nm semiconductor laser were performed with the same total light dose of 30 J/cm2 after the mice were intravenously injection of Rose Bengal for 25 mg/Kg body weight. Laser speckle imaging and microscope were used to monitor blood flow dynamics and vessel constriction during and after V-PDT, respectively. The V-PDT induced vessel damages between different groups were compared. The results show that significant difference in blood vessel damage was found between the lower irradiances (50, 75 and 100 mW/cm2) and higher irradiance (200 mW/cm2), and the blood vessel damage induced by V-PDT is positively correlated with irradiance. This study implies that the optimization of irradiance is required for enhancing V-PDT therapeutic efficiency.
Improving the signal-to-noise ratio in ultrasound-modulated optical tomography by a lock-in amplifier
Lili Zhu, Jingping Wu, Guimin Lin, et al.
With high spatial resolution of ultrasonic location and high sensitivity of optical detection, ultrasound-modulated optical tomography (UOT) is a promising noninvasive biological tissue imaging technology. In biological tissue, the ultrasound-modulated light signals are very weak and are overwhelmed by the strong unmodulated light signals. It is a difficulty and key to efficiently pick out the weak modulated light from strong unmodulated light in UOT. Under the effect of an ultrasonic field, the scattering light intensity presents a periodic variation as the ultrasonic frequency changes. So the modulated light signals would be escape from the high unmodulated light signals, when the modulated light signals and the ultrasonic signal are processed cross correlation operation by a lock-in amplifier and without a chopper. Experimental results indicated that the signal-to-noise ratio of UOT is significantly improved by a lock-in amplifier, and the higher the repetition frequency of pulsed ultrasonic wave, the better the signal-to-noise ratio of UOT.
Synthesis and application of the reduction-sensitive drug delivery system based on water-soluble ZnInAgS quantum dots
Dawei Deng, Rong Zhang, Lingzhi Qu, et al.
High-quality water-soluble quantum dots had been synthesized following the one-step method. Furtherly, the impact factors on the optical properties of quantum dots, which were the feed ratio of S/In and the reflux time, had been concerned emphatically. By changing the reaction parameters, we made the fluorescence emission of ZnInAgS quantum dot tunable from green to orange, and the maximum fluorescence quantum efficiency was up to 30%. Then we modified bovine serum albumin (BSA) on the surface of ZnInAgS quantum dot, which was conjugated with BSA-QDs through the linker of 3'3-dimercapto-diacetate DOX. Finally, the reduction-sensitive drug delivery system based on ZnInAgS quantum dots (QBSSD) was successfully constructed. The resultant QBSSD complex were observed to be significantly stable in aqueous solution. In addition, owing to their cellular reduction responsiveness at the cleavable disulfide linker, the QBSSD complex were able to release DOX rapidly. In vitro drug release and cell level release experiments proved that our QBSSD complexes could make a quick drug release in the environment with GSH. The efficacy experiments showed that our QBSSD complexes exhibited a strong killing effect to cancer cells, and low toxic to normal cells. All the results indicated that the reduction-sensitive drug delivery system was a promising model of administration.
Depth-section imaging of swine kidney by spectrally encoded microscopy
The kidneys are essential regulatory organs whose main function is to regulate the balance of electrolytes in the blood, along with maintaining pH homeostasis. The study of the microscopic structure of the kidney will help identify kidney diseases associated with specific renal histology change. Spectrally encoded microscopy (SEM) is a new reflectance microscopic imaging technique in which a grating is used to illuminate different positions along a line on the sample with different wavelengths, reducing the size of system and imaging time. In this paper, a SEM device is described which is based on a super luminescent diode source and a home-built spectrometer. The lateral resolution was measured by imaging the USAF resolution target. The axial response curve was obtained as a reflect mirror was scanned through the focal plane axially. In order to test the feasibility of using SEM for depth-section imaging of an excised swine kidney tissue, the images of the samples were acquired by scanning the sample at 10 μm per step along the depth direction. Architectural features of the kidney tissue could be clearly visualized in the SEM images, including glomeruli and blood vessels. Results from this study suggest that SEM may be useful for locating regions with probabilities of kidney disease or cancer.
Non-invasive optical detection of HBV based on serum surface-enhanced Raman spectroscopy
Zuci Zheng, Qiwen Wang, Cuncheng Weng, et al.
An optical method of surface-enhanced Raman spectroscopy (SERS) was developed for non-invasive detection of hepatitis B surface virus (HBV). Hepatitis B virus surface antigen (HBsAg) is an established serological marker that is routinely used for the diagnosis of acute or chronic hepatitis B virus(HBV) infection. Utilizing SERS to analyze blood serum for detecting HBV has not been reported in previous literature. SERS measurements were performed on two groups of serum samples: one group for 50 HBV patients and the other group for 50 healthy volunteers. Blood serum samples are collected from healthy control subjects and patients diagnosed with HBV. Furthermore, principal components analysis (PCA) combined with linear discriminant analysis (LDA) were employed to differentiate HBV patients from healthy volunteer and achieved sensitivity of 80.0% and specificity of 74.0%. This exploratory work demonstrates that SERS serum analysis combined with PCA-LDA has tremendous potential for the non-invasive detection of HBV.
Development of a new first-aid biochemical detector
Jingfei Hu, Haiyang Liao, Shilin Su, et al.
The traditional biochemical detector exhibits poor adaptability, inconvenient carrying and slow detection, which can’t meet the needs of first-aid under field condition like natural or man-made disasters etc. Therefore a scheme of first-aid biochemical detector based on MOMES Micro Spectrometer, UV LED and Photodiode was proposed. An optical detection structure combined continuous spectrum sweep with fixed wavelength measurement was designed, which adopted mobile detection optical path consisting of Micro Spectrometer and Halogen Lamp to detect Chloride (Cl-), Creatinine (Cre), Glucose (Glu), Hemoglobin (Hb). The UV LED and Photodiode were designed to detect Potassium (K-), Carbon dioxide (CO2), Sodium (Na+). According to the field diagnosis and treatment requirements, we designed the embedded control hardware circuit and software system, the prototype of first-aid biochemical detector was developed and the clinical trials were conducted. Experimental results show that the sample's absorbance repeatability is less than 2%, the max coefficient of variation (CV) in the batch repeatability test of all 7 biochemical parameters in blood samples is 4.68%, less than the clinical requirements 10%, the correlation coefficient (R2) in the clinical contrast test with AU5800 is almost greater than 0.97. To sum up, the prototype meets the requirements of clinical application.
Retinal image quality and visual stimuli processing by simulation of partial eye cataract
Maris Ozolinsh, Olga Danilenko, Varvara Zavjalova
Visual stimuli were demonstrated on a 4.3’’ mobile phone screen inside a “Virtual Reality” adapter that allowed separation of the left and right eye visual fields. Contrast of the retina image thus can be controlled by the image on the phone screen and parallel to that at appropriate geometry by the AC voltage applied to scattering PDLC cell inside the adapter. Such optical pathway separation allows to demonstrate to both eyes spatially variant images, that after visual binocular fusion acquire their characteristic indications. As visual stimuli we used grey and different color (two opponent components to vision – red-green in L*a*b* color space) spatially periodical stimuli for left and right eyes; and with spatial content that by addition or subtraction resulted as clockwise or counter clockwise slanted Gabor gratings. We performed computer modeling with numerical addition or subtraction of signals similar to processing in brain via stimuli input decomposition in luminance and color opponency components. It revealed the dependence of the perception psychophysical equilibrium point between clockwise or counter clockwise perception of summation on one eye image contrast and color saturation, and on the strength of the retinal aftereffects. Existence of a psychophysical equilibrium point in perception of summation is only in the presence of a prior adaptation to a slanted periodical grating and at the appropriate slant orientation of adaptation grating and/or at appropriate spatial grating pattern phase according to grating nods. Actual observer perception experiments when one eye images were deteriorated by simulated cataract approved the shift of mentioned psychophysical equilibrium point on the degree of artificial cataract. We analyzed also the mobile devices stimuli emission spectra paying attention to areas sensitive to macula pigments absorption spectral maxima and blue areas where the intense irradiation can cause in abnormalities in periodic melatonin regeneration and deviations in regular circadian rhythms. Therefore participants in vision studies using “Virtual Reality” appliances with fixed vision fields and emitting a spike liked spectral bands (on basis of OLED and AMOLED diodes) different from spectra of ambient illuminators should be accordingly warned about potential health risks.
Spectral domain optical coherence tomography with extended depth-of-focus by aperture synthesis
En Bo, Linbo Liu
We developed a spectral domain optical coherence tomography (SD-OCT) with an extended depth-of-focus (DOF) by synthetizing aperture. For a designated Gaussian-shape light source, the lateral resolution was determined by the numerical aperture (NA) of the objective lens and can be approximately maintained over the confocal parameter, which was defined as twice the Rayleigh range. However, the DOF was proportional to the square of the lateral resolution. Consequently, a trade-off existed between the DOF and lateral resolution, and researchers had to weigh and judge which was more important for their research reasonably. In this study, three distinct optical apertures were obtained by imbedding a circular phase spacer in the sample arm. Due to the optical path difference between three distinct apertures caused by the phase spacer, three images were aligned with equal spacing along z-axis vertically. By correcting the optical path difference (OPD) and defocus-induced wavefront curvature, three images with distinct depths were coherently summed together. This system digitally refocused the sample tissue and obtained a brand new image with higher lateral resolution over the confocal parameter when imaging the polystyrene calibration beads.