The Moscone Center
San Francisco, California, United States
28 January - 2 February 2017
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Translational Research Presentations

Biomedical Spectroscopy, Microscopy, and Imaging
(ordered by start date and time)


Molecular imaging of melanin distribution in vivo and quantitative differential diagnosis of human pigmented lesions using label-free harmonic generation biopsy
Paper 10069-4

Author(s):  Chi-Kuang Sun, National Taiwan Univ. (Taiwan), et al.
Conference 10069: Multiphoton Microscopy in the Biomedical Sciences XVII
Session 1: Keynote Session
Date and Time: Sunday, January 29, 2017, 10:30 AM

Melanin is produced by melanocytes in the epidermal basal layer and is the major determinant of skin color. Clinical molecular imaging of melanin with a sub-cellular resolution is not only critical for hypopigmentary/hyperpigmentary skin disease treatment monitoring, but also for differential diagnosis of pigmented skin tumors. In this talk, by presenting all different physics and clinical trial studies, we demonstrate that harmonic generation microscopy is the only proven technique to meet the above-mentioned clinical need for noninvasive label-free 3D imaging with a histopathological accuracy while high penetration, no photodamage, and high resolution can all be satisfied at the same time.


Structural determinations in ovarian cancer via pixel based SHG polarization analyses
Paper 10069-6

Author(s):  Paul J. Campagnola, Univ. of Wisconsin-Madison (United States), et al.
Conference 10069: Multiphoton Microscopy in the Biomedical Sciences XVII
Session 2: Second/Third Harmonic Generation I
Date and Time: Sunday, January 29, 2017, 11:20 AM

We use three SHG polarization metrics to probe ECM remodeling in human ovarian cancer which determine i) the helical pitch angle via the single axis molecular model, ii) dipole alignment via anisotropy, and iii) chirality via SHG circular dichroism. These methods differentiate Col I and Col III in fibrillar gel models and also human ovarian tissues (normal and tumors) due to differences in the α-helix angle of the isoforms and are consistent with immunofluorescence and gene expression measurements. These SHG polarization methods provide sub-resolution structural information not otherwise possible and can serve as label-free biomarkers for ovarian and other cancers.


Single cell genomic quantification by non-fluorescence nonlinear microscopy
Paper 10071-20

Author(s):  Jing Liu, South Dakota School of Mines and Technology (United States), et al.
Conference 10071: Single Molecule Spectroscopy and Superresolution Imaging X
Session 6: Biological Applications of Single Molecule Detection Techniques
Date and Time: Sunday, January 29, 2017, 11:30 AM


A compact, robust, high throughput digital holographic microscope with high sensitivity
Paper 10074-13

Author(s):  Manuel Bedrossian, California Institute of Technology (United States), et al.
Conference 10074: Quantitative Phase Imaging III
Session 2: QPI Methodologies II
Date and Time: Sunday, January 29, 2017, 3:40 PM

We present the design of a compact and robust off-axis digital holographic microscope (DHM) and results from imaging samples spiked with 0-108 bacteria/mL. Guidelines for imaging times required to achieve different levels of detection sensitivity are presented, and an upper limit to bacterial counting is also shown, illustrating the sample turbidity at which hologram reconstruction is no longer possible. The instrument is an ‘all-in-one’ fully autonomous off-axis DHM with its own electronics and sample loading and unloading abilities, making it a promising instrument for field diagnostics and high-throughput bacterial screening of liquid samples.


Moxifloxacin: Clinically compatible contrast agent for multiphoton imaging
Paper 10069-80

Author(s):  Taejun Wang, Pohang Univ. of Science and Technology (Korea, Republic of), et al.
Conference 10069: Multiphoton Microscopy in the Biomedical Sciences XVII
Session PSun: Posters-Sunday
Date and Time: Sunday, January 29, 2017, 5:30 PM

Fluorescence-based cellular-level imaging of human tissues has been challenging due to concerns of cytotoxicity of labeling agents or photodamage by high laser power. Herein, we describe the applications of moxifloxacin, an FDA approved antibiotic, as a cell labeling agent for multiphoton microscopy (MPM). Quick penetration and high intracellular concentration of moxifloxacin allowed MPM visualization of cellular morphology and dynamics within tissues at significantly reduced laser power compared to autofluorescence-based imaging.


Transient absorption imaging of glycated hemoglobin in blood of diabetes patients
Paper 10069-82

Author(s):  Pu-Ting Dong, Purdue Univ. (United States), et al.
Conference 10069: Multiphoton Microscopy in the Biomedical Sciences XVII
Session PSun: Posters-Sunday
Date and Time: Sunday, January 29, 2017, 5:30 PM


Multimodal optical imaging database from tumour brain human tissue: endogenous fluorescence from glioma, metastasis and control tissues
Paper 10069-94

Author(s):  D. Abi-Haidar, Institut National de Physique Nucléaire et de Physique des Particules (France), et al.
Conference 10069: Multiphoton Microscopy in the Biomedical Sciences XVII
Session PSun: Posters-Sunday
Date and Time: Sunday, January 29, 2017, 5:30 PM

Surgical resection, whenever feasible, is the first-line therapy to treat central nervous system tumours. The identification of the tumour margins, including glioma, metastasis and meningioma is difficult intraoperatively. We aim to construct a database of multimodal optical imaging information from fresh and fixed human samples. Our team used four different contrasts. The results of multimodal optical analysis on human biopsies were compared to the gold standard histopathology.


Imaging of epithelial-mesenchymal transition process of live gastric cancer cells with spectral focusing based hyperspectral stimulated Raman scattering
Paper 10069-96

Author(s):  Zi Wang, National Univ. of Singapore (Singapore), et al.
Conference 10069: Multiphoton Microscopy in the Biomedical Sciences XVII
Session PSun: Posters-Sunday
Date and Time: Sunday, January 29, 2017, 5:30 PM

In this paper, we report the development of a spectral focusing based hyperspectral stimulated Raman scattering (SRS) imaging technique for label-free imaging of the biochemical/biomolecular and morphological changes in live gastric cancer cells during the epithelial-mesenchymal transition (EMT) process. The developed SRS imaging technique can cover the 2800-3100 cm-1 region in ~10 secs by scanning the time delay between the chirped pump and Stokes beams. The distributions of proteins, lipids and DNA can be extracted from the hyperspectral SRS images using multivariate curve resolution (MCR) method. Our results show that the hyperspectral SRS imaging technique is a powerful label-free imaging method for visualizing the morphological and biological/biochemical change during the EMT process and other biological activities of live cells


Oxidatively generated base damage to DNA in aqueous solutions by femtosecond laser-induced low density plasma multi-channels controlled with a spatial light modulator
Paper 10075-27

Author(s):  Hakim Belmouaddine, Univ. de Sherbrooke (Canada), et al.
Conference 10075: Biophysics, Biology and Biophotonics II: the Crossroads
Session PSun: Posters-Sunday
Date and Time: Sunday, January 29, 2017, 5:30 PM

To better understand the radiation chemistry following from the generation of low density plasma in systems of biological interest, we encode a micro-lens array into a spatial light modulator to harness the multi-filamentation of powerful near-infrared femtosecond laser pulses in aqueous solutions of DNA. The present method of irradiation allows the generation of a programmable matrix of laser-induced low density plasma channels in water. From the analysis of the laser-induced DNA damages, we conclude that each plasma channel behaves as an independent intense micro beam of ionizing radiation capable of yielding complex DNA damage.


Portable SERS sensor for malachite green and other small dye molecules
Paper 10072-1

Author(s):  Jingting Li, Univ. of Houston (Uganda), et al.
Conference 10072: Optical Diagnostics and Sensing XVII: Toward Point-of-Care Diagnostics
Session 1: Optical Biosensing Using Surface Enhanced Raman Spectroscopy
Date and Time: Monday, January 30, 2017, 8:20 AM

Sensitive detection of specific chemicals on site can be extremely powerful in many fields. Owing to its molecular fingerprinting capability, surface-enhanced Raman scattering has been one of the technological contenders. In this paper, we describe the novel use of DNA topological nanostructure on nanoporous gold nanoparticle (NPG-NP) array chip for chemical sensing. . NPG-NP features large surface area and high-density plasmonic field enhancement known as “hot-spots”. Hence, NPG-NP array chip has found many applications in nanoplasmonic sensor development. This technique can provide novel label-free molecular sensing capability with high sensitivity and specificity.


New mononuclear leukocyte-like populations within the granulocyte scatter gate detected by flow cytometry
Paper 10068-3

Author(s):  Attila Tárnok, Univ. Leipzig (Germany), et al.
Conference 10068: Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XV
Session 1: Cytomics and Histomics
Date and Time: Monday, January 30, 2017, 9:00 AM

Granulocytes are central in innate immunity. We detected previously unknown lymphocyte- (LL) and monocyte-like (ML) cells in the granulocyte scatter gate. 905 healthy adults from the LIFE study were immunophenotyped by multiplexed flow cytometry (420 men; age:56.5±14.0 years; 485 women:56.7±13.6 y). Values compared statistically: men vs women, young (18-49 y) vs elderly (50-81 y.) men, and women. Four CD45+, SSCmid-high cell types were detected: LL1=CD3+,CD4+,CD8++,CD16/56+,CD38+,HLA-DR+ LL2=CD3+,CD4low,CD8+,CD38low LL3=CD3+,CD4+,CD8- ML1=CD3-,CD4low,CD14+,CD38+ LL2 and ML1 counts increased in men (p<0.04); LL2 in women was age dependent (p<0.001). New cell types with neutrophil scatter characteristics are reported. Characterization in health and diseases is planned.


Functional imaging of live Zebrafish using fluorescence lifetime optical projection tomography
Paper 10068-7

Author(s):  Natalie Andrews, Imperial College London (United Kingdom), et al.
Conference 10068: Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XV
Session 2: Functional Imaging I
Date and Time: Monday, January 30, 2017, 10:10 AM

Current microscopy techniques are not optimal to image fluorescence in whole live animals. We present fluorescence lifetime optical projection tomography (FLIM OPT) applied to functional imaging in live transgenic zebrafish expressing a Caspase 3 Förster Resonance Energy Transfer (FRET) biosensor. We can detect changes in Caspase 3 activity in embryo Tg(Ubi:Caspase3biosensor) zebrafish after induction of apoptosis by gamma irradiation. Though development of compressive sensing and multiplexed imaging with two imaging arms we have applied OPT and FLIM OPT to adult zebrafish, enabling us to quickly acquire datasets so the fish can be recovered and imaged longitudinally.


Application of broadband coherent Raman imaging to histopathology
Paper 10069-25

Author(s):  Marcus T. Cicerone, National Institute of Standards and Technology (United States), et al.
Conference 10069: Multiphoton Microscopy in the Biomedical Sciences XVII
Session 7: Biomedical Applications of Coherent Raman II
Date and Time: Monday, January 30, 2017, 10:30 AM

We will report on application of broadband coherent anti-Stokes Raman scattering microscopy1 to chemical mapping and characterization of resected prostate sections. While incidence of prostate cancer is very high, only a small fraction of prostate tumors will progress to advanced, metastatic disease and become dangerous, but prostatectomy and follow-on treatment have many undesirable potential side effects. Thus, it is important to predict which tumors will progress and which should be removed, but there is currently no highly reliable way to make such predictions. We will present a retrospective coherent Raman imaging study resected prostate sections focusing on locating tissue regions that present the highest diagnostic value with respect to lethal vs indolent disease. We intend that this provide a guide to optimal spectral sampling of these tissues to address this important clinical problem.


Label-free biomolecular characterization of human breast cancer tissue with stimulated Raman scattering (SRS) spectral imaging
Paper 10069-27

Author(s):  Fa-Ke F. Lu, Brigham and Women's Hospital, Harvard Medical School (United States), et al.
Conference 10069: Multiphoton Microscopy in the Biomedical Sciences XVII
Session 7: Biomedical Applications of Coherent Raman II
Date and Time: Monday, January 30, 2017, 11:10 AM

SRS microscopy is used to characterize the biomolecules in breast cancer tissue. Human breast cancer specimens at the tumor core, tumor margin and normal area (5 cm away from the tumor) from surgical cases will be imaged with SRS at multiple Raman shifts, including the peaks for lipid, protein, blood (absorption), collagen, microcalcification (calcium phosphates and calcium oxalate) and carotenoids. Most of these Raman shifts have relatively strong Raman cross sections, which ensures high-quality and fast imaging. This proof-of-principle study is sought to demonstrate the feasibility and potential of SRS imaging for ambient diagnosis and surgical guidance of breast cancer.


The light at the service of medicine: optical sensing beside the patient's bed
Paper 10072-10

Author(s):  Francesco Baldini, Istituto di Fisica Applicata "Nello Carrara" (Italy), et al.
Conference 10072: Optical Diagnostics and Sensing XVII: Toward Point-of-Care Diagnostics
Session 3: Optical Point of Care Sensing: At The Bedside
Date and Time: Monday, January 30, 2017, 1:20 PM

Optical biosensors and chemosensors can definitely play a fundamental role in the development of POCT devices also in the case of invasive measurements. The activity developed at the Institute of Applied Physics in this field in strict collaboration with physicians is described with particular attention to the measurement of bile-containing refluxes in the gastroesophageal apparatus in non- hospitalized patients, to the detection of gastric carbon dioxide in intensive care patients, to the simultaneous measurement of sepsis biomarkers in serum samples and to the measurements of immuno-suppressants in transplanted patients.


Novel fluorescence-based POCT platform for therapeutic drug monitoring in transplanted patients
Paper 10072-11

Author(s):  Francesco Baldini, Istituto di Fisica Applicata "Nello Carrara" (Italy), et al.
Conference 10072: Optical Diagnostics and Sensing XVII: Toward Point-of-Care Diagnostics
Session 3: Optical Point of Care Sensing: At The Bedside
Date and Time: Monday, January 30, 2017, 1:40 PM

One of the most challenging objectives in the clinical treatment of transplanted patients, is the therapeutic drug monitoring (TDM) of immunosuppressants which, while protecting the transplanted organ from rejection, exhibit a very narrow therapeutic window and a high degree of variability. The key to improve long-term outcome following the transplantation is the selection of the correct immunosuppressive regime to minimize toxicity and the risk of adverse events, while maintaining immunosuppressive efficacy. A novel TDM point of care testing (POCT) optical device for the detection of immunosuppressants was designed and tested, with the body interface constituted by an intravascular microdialysis catheter (MicroEye®), which provides the dialysate as clinical sample. An optical biochip with 10 microchannels (length: 14 mm, width: 500 μm, depth: 140 μm), based on total internal reflection fluorescence (TIRF), enables the frequent measurement of immunosuppressants. Heterogeneous competitive immunoassays for the detection of mycophenolic acid, tacrolimus and cyclosporine A are implemented on the different microchannels, with the derivative of the immunosuppressants immobilised on the bottom part of the micro-channels. Detection limits of 0.05 ng mL-1 and 0.78 ng mL-1 were achieved for tacrolimus and mycophenolic acid, respectively, while, for cyclosporine A, a binding inhibition rate of 62% was achieved with 10 ng mL-1 of the analyte. In order to achieve optimisation of the performances, the detection antibodies of the different immunosuppressants have been immobilised onto magnetic nanoparticles, and suitable magnetic traps interfaced with the optical chip are used to drive magnetically the implementation of the assay.


Monitoring of live cell cultures during apoptosis by phase imaging and Raman spectroscopy
Paper 10074-29

Author(s):  Alexander T. Khmaladze, Univ. at Albany (United States), et al.
Conference 10074: Quantitative Phase Imaging III
Session 5: QPI of Cells and Tissues I
Date and Time: Monday, January 30, 2017, 2:55 PM

Non-invasive label-free live cell measurements are an important tool in biomedical research. We present a combined digital holography/Raman spectroscopy technique for cell monitoring. Digital holographic microscopy records an interference pattern between object and reference waves, so that a 3D image is obtained. This technique yields information about cell cycle and cell death mechanisms, since these processes are correlated with individual cell volume and shape. Raman spectroscopy contains rotational and vibrational molecular transitions, as well as intermolecular vibrations; therefore, complementary information about cell content can be gained. We analyze and compare cell culture data obtained by the two methods.


Toward unstained cytology and complete blood counts at the point of care
Paper 10072-15

Author(s):  Andres F. Zuluaga, Clearview App, Inc. (United States), et al.
Conference 10072: Optical Diagnostics and Sensing XVII: Toward Point-of-Care Diagnostics
Session 4: Optical Point of Care Technologies For Sensing: In The Field
Date and Time: Monday, January 30, 2017, 3:30 PM

There is a critical unmet need for an instrument to perform complete blood counts (CBCs) at the point of care. There is currently no product in the US that can perform these bedside. We’ve developed systems capable of tomographic images with sub-cellular resolution with consumer-grade broadband (LED) sources and CMOS detectors suitable for POC implementation of CBC tests. We‘ll show results imaging and differentiating unstained blood cells using supercontinuum lasers and LEDs as sources and CMOS camera sensors, and lay out the follow up steps needed: image segmentation, analysis, and classification, advancing toward bedside CBCs that don't require CLIA-certified laboratories.


Field performance of a low-cost and fully-automated blood counting system operated by trained and untrained users
Paper 10072-16

Author(s):  Dengling Xie, Univ. of Science and Technology of China (China), et al.
Conference 10072: Optical Diagnostics and Sensing XVII: Toward Point-of-Care Diagnostics
Session 4: Optical Point of Care Technologies For Sensing: In The Field
Date and Time: Monday, January 30, 2017, 3:50 PM

We report here a method to count red blood cells, white blood cells and platelets through a low-cost and fully-automated blood counting system, comparing its performance between trained and untrained users. It uses a single-step sample preparation method and autofocus to reduce dependence on highly trained technicians. Results on human and child blood demonstrate close agreement with a clinical standard and indicate that the device could have application in low-resource areas where central hematology laboratories are not accessible.


DotLens smartphone microscopy for biological and biomedical applications
Paper 10072-18

Author(s):  Wei-Chuan Shih, Univ. of Houston (United States), et al.
Conference 10072: Optical Diagnostics and Sensing XVII: Toward Point-of-Care Diagnostics
Session 4: Optical Point of Care Technologies For Sensing: In The Field
Date and Time: Monday, January 30, 2017, 4:30 PM

Recent advances in inkjet-printed optics have created a new class of lens fabrication technique. Dubbed DotLens, a single of which weighs less than 50 mg and occupies a volume less than 50 μL. DotLens can be attached onto any smartphone camera akin to a contact lens, and turn the smartphones into a microscope. In this paper, we show recent results from images collected from pathology tissue slides with cancer features. In addition, we show performance in cytological analysis of blood smear. This tool has empowered Citizen Science investigators to collect microscopic images from various interesting objects.


Multispectral imaging based on a smartphone with an external C-MOS camera for detection of seborrheic dermatitis of a scalp
Paper 10068-65

Author(s):  Manjae Kim, DGIST (Korea, Republic of), et al.
Conference 10068: Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XV
Session PMon: Posters-Monday
Date and Time: Monday, January 30, 2017, 5:30 PM

The key thing of the manuscript is the self-diagnosis of seborrheic dermatitis of a scalp at home by using multispectral imaging based on a smartphone: Because of that, we integrated a very small CMOS camera, which is typically utilized in a smartphone, with a LED modules. After that, it is directly connected to a smartphone by a OTG cable for self-diagnosis of seborrheic dermatitis of a scalp to control the device and perform spectral imaging and analysis. In fact, it was found that it is very difficult to self-diagnose seborrheic dermatitis of a scalp using our previous version of the smartphone-based multispectral imaging system. Note that the size of the developed device is less than 77 X 33 X 16 mm. Therefore, it is very compact and appropriated as a handheld device to carry out multispectral imaging of seborrheic dermatitis of a scalp for self-diagnosis of the seborrheic dermatitis using a smartphone integrated with the device. For the spectral classification and analysis of the acquired images, a smartphone App, we developed, was utilized. 3) Finally, by using the system, we performed multispectral imaging of seborrheic dermatitis regions and normal regions of a scalp and moreover discriminated between seborrheic dermatitis regions and acne or psoriasis, which looks similar to the seborrheic dermatitis regions, in the scalp. Therefore, we demonstrate that the multispectral imaging based on a smartphone have the great potentials for self-diagnosis of seborrheic dermatitis of the scalp at home.


Quantitative phase imaging of platelet: assessment of cell morphology and function
Paper 10068-70

Author(s):  Irina Vasilenko, Maimonides State Classical Academy (Russian Federation), et al.
Conference 10068: Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XV
Session PMon: Posters-Monday
Date and Time: Monday, January 30, 2017, 5:30 PM

Preliminary work has studied the accompanying shape change, wherein platelets undergo a major cytoskeletal rearrangement where the cell body swells up and then flattens out to form extensions called lamellipodia and filopodia, but the timeline of shape change has not been quantitatively evaluated. The different types of protrusions have different physiological roles: filopodia connect individual platelets to each other, whereas lamellipodia help the platelets to cover the wounded area. The use of the computer phase-inteference microscope could be an easy and fast method to check the changed platelet activation and to follow a possible pharmacological therapy to reduce this phenomenon.


Novel system for measuring giant spectral images and its application for cancer detection
Paper 10068-21

Author(s):  Yuval Garini, bar Ilan University (Israel), et al.
Conference 10068: Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XV
Session 6: Image Analysis
Date and Time: Tuesday, January 31, 2017, 8:00 AM

Spectral imaging is an important method that is used for various applications, but measuring very large spectral images is a challenge that so far was not achieved. We present a novel system for scanning very large spectral images of microscopy samples in a rather short time. The system captures the information while the sample is continuously being scanned on the fly. It therefore breaks the size and speed limits that resulted from existing spectral imaging methods. The spectral separation is achieved through Fourier spectroscopy by using an interferometer mounted along the optical axis (no moving parts). We will describe the system and its use for pathological samples and cancer detection.


Feeling for cell function: Mechanical phenotyping at 1,000 cells/sec
Paper 10076-21

Author(s):  Jochen R. Guck, TU Dresden (Germany), et al.
Conference 10076: High-Speed Biomedical Imaging and Spectroscopy II: Toward Big Data Instrumentation and Management
Session 5: High-throughput Imaging I
Date and Time: Tuesday, January 31, 2017, 8:30 AM

The mechanical properties of cells can be seen as a label-free, inherent marker of biological function and disease, but a simple and convenient measurement technique with sufficient throughput has been lacking. To address this need, we have introduced real-time deformability cytometry (RT-DC) for the continuous mechanical single-cell characterization with analysis rates up to 1,000 cells/s. Our results indicate that cell mechanics can define cell function, can be used as an inherent cell marker and could serve as target for novel therapies. Mechanical phenotyping adds a new functional, marker-free dimension to flow cytometry with diverse applications in biology, biotechnology and medicine.


Use of Gabor filters and deep networks in the segmentation of retinal vessel morphology
Paper 10068-25

Author(s):  Henry Leopold, Univ. of Waterloo (Canada), et al.
Conference 10068: Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XV
Session 6: Image Analysis
Date and Time: Tuesday, January 31, 2017, 9:20 AM

The first step of almost all forms of fundus analysis begins with the segmentation and subtraction of retinal vasculature, while analysis of that same structure can aid in the diagnosis of retinal and cardiovascular conditions, such as diabetes or stroke. Accurate feature extraction from retinal fundus images is essential for physicians, optometrists and ophthalmologists in the care and treatment of their patients. This work explores the use of Computer aided retinal vessel segmentation as a precursor for the early detection of conditions that present abnormalities within the vasculature, such as coronary heart disease, stroke and Diabetes.


Effect of sample bias on accuracy of a three-part leukocyte differential test when using AO-induced fluorescence and colorimetric features
Paper 10072-23

Author(s):  Amy J. Powless, Univ. of Arkansas (United States), et al.
Conference 10072: Optical Diagnostics and Sensing XVII: Toward Point-of-Care Diagnostics
Session 5: Fluorescent Assays for Optical Sensing and Imaging
Date and Time: Tuesday, January 31, 2017, 9:40 AM

Many point-of-care (POC) systems performing a three-part leukocyte differential use acridine orange (AO) as a fluorescent contrast agent due to its colorimetric features, however, time-dependent red shift can limit image acquisition and thus sample size which could cause test inaccuracy. We have developed an automated differential algorithm and used Bland-Altman analysis to quantify accuracy. We have investigated the effect of sample bias on test accuracy using AO with our POC device that can count an average of 200 cells per field-of-view (FOV). Currently, we have produced results within +/-15% of a clinical system using six FOVs.


Improved cancer risk stratification and diagnosis via quantitative phase microscopy
Paper 10074-40

Author(s):  Yang Liu, Univ. of Pittsburgh (United States), et al.
Conference 10074: Quantitative Phase Imaging III
Session 7: QPI Clinical Applications
Date and Time: Tuesday, January 31, 2017, 10:40 AM

We present the use of quantitative phase microscopy to assess nanoscale nuclear architecture that is not easily identifiable by conventional pathology, to improve cancer diagnosis of cytopathology and early prediction of cancer progression.


Opportunities of QPI in the epigenetic diagnostics and assessment of therapeutic efficacy
Paper 10074-41

Author(s):  Irina Vasilenko, Maimonides State Classicial Academy (Russian Federation), et al.
Conference 10074: Quantitative Phase Imaging III
Session 7: QPI Clinical Applications
Date and Time: Tuesday, January 31, 2017, 11:10 AM

Analysis of the chromosome areas nuclear distribution and location of some areas of lymphocyte chromatin in peripheral blood was carried out in 75 patients with verified multiple sclerosis. The control group included 20 healthy volunteers. Investigation was performed in the real time using apparatus-software complex “Bioni” (Moscow) for clinical and laboratory diagnostics. Software packet enables receiving phase portrait of the cell nucleus and its fragments, file editing and inversion, files subtraction, fluctuation mapping, etc. Analysis of the peculiarities of chromatin structure as biomarker may be applied in diagnostics of a series of diseases. An important aspect of the development of this scientific direction includes modern technologies of visualization and digital processing of biological structure images at different levels using QPI.


In vivo multiphoton imaging and quantification of cytoplasmic and nuclear metabolism in the hepatobiliary system
Paper 10069-54

Author(s):  Chen-Yuan Dong, National Taiwan Univ. (Taiwan), et al.
Conference 10069: Multiphoton Microscopy in the Biomedical Sciences XVII
Session 12: Metabolism-NADH/FAD/Tryptophan I
Date and Time: Tuesday, January 31, 2017, 11:50 AM


Optical monitoring of spinal cord hemodynamics
Paper 10072-28

Author(s):  Babak Shadgan, The Univ. of British Columbia (Canada), et al.
Conference 10072: Optical Diagnostics and Sensing XVII: Toward Point-of-Care Diagnostics
Session 7: Spectroscopy Experimental and Modeling Toward Perfusion and Oxygenation Monitoring
Date and Time: Tuesday, January 31, 2017, 1:20 PM

Spinal cord (SC) ischemia and hypoxia are important contributors to secondary damage after spinal cord injury (SCI). To mitigate these secondary processes and improve neurologic outcome, current clinical practice guidelines recommend aggressive maintenance of SC perfusion and oxygenation. Such hemodynamic management, however, is currently carried out without any real-time measures of SC perfusion and oxygenation. Such information would be extremely valuable to clinicians managing SCI patients in the intensive care setting. This study examined the feasibility and sensitivity of a custom-made miniaturized near-infrared spectroscopy sensor to monitor SC hemodynamics and oxygenation in a pig model of SCI.


In vivo preclinical verification of a multimodal diffuse reflectance and correlation spectroscopy system for sensing tissue perfusion
Paper 10072-29

Author(s):  Julia M. Pakela, Univ. of Michigan (United States), et al.
Conference 10072: Optical Diagnostics and Sensing XVII: Toward Point-of-Care Diagnostics
Session 7: Spectroscopy Experimental and Modeling Toward Perfusion and Oxygenation Monitoring
Date and Time: Tuesday, January 31, 2017, 1:40 PM

In reconstructive surgery, impeded blood flow in tissue grafts due to arterial or venous clots can rapidly result in graft failures. Thus, the ability to detect changes in tissue perfusion in microvascular free grafts is critical. We report progress on in vivo pre-clinical testing of a compact, multimodal, fiber-based diffuse correlation and reflectance spectroscopy system designed to quantitatively monitor blood perfusion in surgically-grafted free flaps in a porcine model. We describe device sensitivity to incremental changes in blood flow and discuss the prospects for continuous perfusion monitoring in future clinical translational studies.


Effects of anti-cancer drug doxorubicin on endogenous biomarkers NAD(P)H, FAD & Trp in prostate cancer cells: a FLIM Study
Paper 10069-56

Author(s):  Horst K. Wallrabe, Univ. of Virginia (United States), et al.
Conference 10069: Multiphoton Microscopy in the Biomedical Sciences XVII
Session 13: Metabolism-NADH/FAD/Tryptophan II
Date and Time: Tuesday, January 31, 2017, 1:45 PM

Fluorescence Lifetime Imaging (FLIM) can be used to identify metabolic changes during cancer progression and upon anti-cancer drug treatment. Prostate cancer (PCa) is one of the leading cancers in men in the USA. Mitochondrial dysfunction and defective OXPHOS (oxidative phosphorylation) activity has been reported in cancer. The anti-cancer drug doxorubicin has been shown to induce apoptosis. Induction of cell death by correction of mitochondrial activity is a promising strategy of cancer treatment. This research focusses on understanding the changes in NAD(P)H, FAD and Trp in mitochondria of LNCaP PCa cells upon treatment with doxorubicin using our 3-channel FLIM-FRET approach and by quantitative analysis of cellular metabolism. Live cell FLIM-FRET measurements on LNCaP cells were done on Zeiss 780 2p confocal microscope coupled with B&H TCSPC FLIM board. The same FoV (fields of view) were imaged both before treatment and in 15 minute intervals after adding 0.5 µM doxorubicin up to 60 min when the cells started to die. Increase in NAD(P)H τm, increase in NAD(P)H enzyme bound fraction, decrease in FAD enzyme bound and reduction in the NAD(P)H/FAD redox ratio was seen upon doxorubicin treatment. Increase in Trp-E% was also seen upon doxorubicin treatment which corresponds to the increase in the NAD(P)H enzyme bound fraction. Our results are very encouraging and demonstrate that FLIM-FRET has the capability to detect early changes associated with the correction in mitochondrial/OXPHOS activity and induction of apoptosis upon treatment with doxorubicin in PCa cells.


Image-guided single cell extraction from mouse calvarium bone marrow in vivo
Paper 10068-28

Author(s):  Christa Haase, Wellman Ctr. for Photomedicine (United States), et al.
Conference 10068: Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XV
Session 8: Instrumentation
Date and Time: Tuesday, January 31, 2017, 2:00 PM

We have developed an image-guided technique for isolating specific acute myeloid leukemia cells from the mouse calvarium bone marrow (BM) in vivo. Accessing the BM in a minimally invasive manner is achieved by plasma-mediated laser bone ablation using a frequency-doubled fiber laser source focused through a high-NA objective lens. Target cells are aspirated with a micropipette and sorted onto a well-plate filled with lysis buffer, which facilitates subsequent single-cell RNA sequencing. This technique opens up the possibility of spatially resolved, image-guided, single-cell transcriptomics of leukemic cells as a function of disease progression.


Quantitative metabolic microendoscopy within a living organism based on two-photon excited endogenous molecular imaging of intracellular NADH and FAD
Paper 10069-58

Author(s):  Pierre Leclerc, XLIM Institut de Recherche (France), et al.
Conference 10069: Multiphoton Microscopy in the Biomedical Sciences XVII
Session 13: Metabolism-NADH/FAD/Tryptophan II
Date and Time: Tuesday, January 31, 2017, 2:20 PM

Multiphoton microscopy is a cutting edge imaging modality leading to remarkable step forward in biology but also in the clinical field. To use it at its full potential and at the very heart of clinical practice, there has been several development of fiber-based micro-endoscope. We demonstrate, for the first time, that it is possible for a multiphoton endoscope, to access and assess fluorescence of weak intrinsic molecule such as intracellular NADH and FAD, thus showing that it is possible to monitor cellular metabolism, in real time, in vivo in situ, without staining, minimally invasively within a living organism.


Pump-probe microscopy of respiratory chain pigments: towards non-fluorescent label-free metabolic imaging
Paper 10069-60

Author(s):  Jesse W. Wilson, Colorado State Univ. (United States), et al.
Conference 10069: Multiphoton Microscopy in the Biomedical Sciences XVII
Session 13: Metabolism-NADH/FAD/Tryptophan II
Date and Time: Tuesday, January 31, 2017, 2:50 PM

Motivated by the success of pump-probe microscopy of hemoglobin, we seek to extend the technique to the cytochromes, with the goal of dissecting respiratory chain function of individual cells in live tissue. Previous transient absorption spectroscopy studies indicate the cytochrome c redox states are differentiable for visible pump-probe pulses. A microscopy technique capable of distinguishing between the redox states of cytochromes will be a valuable addition to respiratory chain imaging, which is currently limited to molecules that fluoresce, such as NAD(P)H and FAD+. Initial imaging results on brown adipose tissue and heart muscle show differences in the averaged pump-probe response.


Single-cell analysis of radiotracers' uptake by fluorescence microscopy: direct and droplet approach
Paper 10068-32

Author(s):  Maria Elena Gallina, Stanford Univ. (United States), et al.
Conference 10068: Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XV
Session 8: Instrumentation
Date and Time: Tuesday, January 31, 2017, 4:10 PM

Recently, radioluminescence microscopy extended the use of radionuclides to single-cell studies. Here we propose a new single-cell radioisotopic assay that improves throughput while adding sorting capabilities. The new method uses fluorescence-based sensors for revealing single-cell interactions with molecular radiomarkers. This study focuses on comparing two methods: the direct approach, where the sensor is incorporated in the cytoplasm, and the droplet approach, where it is co-encapsulated with radiolabeled single-cells in droplets. Both approaches successfully activated the fluorescence signal following cellular uptake of radiotracers. While the direct approach offers higher resolution, the droplet approach is more suitable for studying heterogeneous populations.


Robust phase unwrapping for phase images in Fourier domain Doppler optical coherence tomography
Paper 10070-52

Author(s):  Yong Huang, Beijing Institute of Technology (China), et al.
Conference 10070: Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIV
Session 11: Quantitative Imaging
Date and Time: Wednesday, February 1, 2017, 3:00 PM


Important Dates

Abstracts Due
17 July 2017

Author Notification
25 September 2017

Manuscripts Due
See Individual Conferences


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Journal of Biomedical Optics

Journal of Biomedical OpticsPublishes peer-reviewed papers that utilize modern optical technology for improved health care and biomedical research.