Conference 11654

High-Speed Biomedical Imaging and Spectroscopy VI

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  • Welcome and Introduction
  • BiOS Hot Topics
  • 1: High-speed Photoacoustic Imaging
  • 2: High-speed Multiphoton Imaging I
  • 3: High-speed Multiphoton Imaging II
  • 4: Emerging Technologies I
  • 5: High-Speed Super-Resolution Imaging
  • 6: Imaging Flow Cytometry
  • 7: Emerging Technologies II
  • 8: Deep Learning
  • Poster Session
  • Front Matter: Volume 11654
Session R: Welcome and Introduction
11654-800
Author(s): Keisuke Goda, The Univ. of Tokyo (Japan); Kevin K. Tsia, The Univ. of Hong Kong (Hong Kong, China)
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Introduction to SPIE Photonics West BiOS conference 11654: High-Speed Biomedical Imaging and Spectroscopy VI
Session LIVE: BiOS Hot Topics
11618-700
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Every year, attendees look forward to Saturday Night Hot Topics, an evening spent hearing highly engaged, world renowned speakers reveal the latest innovations in their areas of expertise. Don't miss this year's outstanding list of speakers.
11648-601
Author(s): Enrico Gratton, Univ. of California, Irvine (United States)
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11648-600
Author(s): Kevin K. Tsia, The Univ. of Hong Kong (Hong Kong, China)
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11642-600
Author(s): Chulhong Kim, Pohang Univ. of Science and Technology (Korea, Republic of)
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Session 1: High-speed Photoacoustic Imaging
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Author(s): Junjie Yao, Duke Univ. (United States)
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Optical-resolution photoacoustic microscopy (OR-PAM) has become a popular tool in small-animal studies. However, previous OR-PAM techniques variously lacked a high imaging speed, a high spatial resolution, and/or a large field of view. Here we report a high-speed OR-PAM system using an innovative water-immersible polygon-mirror scanner, which has achieved a cross-sectional frame rate of as high as 2400 Hz over a 12-mm scanning range. Using this polygon-scanner-based OR-PAM system, we have performed various studies on mouse models with stroke and cardiac arrests. We expect that the new OR-PAM system will become a powerful tool for imaging hemodynamics and neuronal functions.
11654-27
Author(s): Song Hu, Washington Univ. in St. Louis (United States)
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Enabling simultaneous imaging of microvascular blood perfusion, oxygenation and flow and high-resolution mapping of tissue oxygen extraction and metabolism in vivo, multi-parametric photoacoustic microscopy (PAM) has opened a new window to reveal the hemodynamic and metabolic insights underlying a wide spectrum of physiological and pathological processes. Although promising, the application and dissemination of this enabling technique have been hindered by the relatively low speed, due to important limitations in image acquisition and data analysis. This talk will focus on our recent efforts in pushing the envelope of multi-parametric PAM towards high-resolution wide-field functional and metabolic imaging in real time through a hardware-software combined approach.
11654-28
Author(s): Jongbeom Kim, Jin Young Kim, Chulhong Kim, Pohang Univ. of Science and Technology (Korea, Republic of)
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Here, we introduce a high-speed super-resolution localization optical-resolution photoacoustic microscopy (OR-PAM) that overcomes limited temporal/spatial resolutions. First, we equipped a galvanometer scanner to improve temporal resolution. Our system achieves a wide scanning range and a high signal-to-noise ratio as well as the B-mode imaging speed of 500 Hz. Furthermore, we acquired super-resolved images in vivo using an agent-free localization technology based on the fast scanning speed. These results show that our OR-PAM can be used various fields, including neurology, oncology, and pathology.
11654-29
Author(s): Alessandro Zunino, Istituto Italiano di Tecnologia (Italy), Univ. degli Studi di Genova (Italy); Francesco Garzella, Istituto Italiano di Tecnologia (Italy), Univ. degli Studi di Parma (Italy); Peter Saggau, Istituto Italiano di Tecnologia (Italy), Baylor College of Medicine (United States); Paolo Bianchini, Istituto Italiano di Tecnologia (Italy); Alberto Diaspro, Istituto Italiano di Tecnologia (Italy), Univ. degli Studi di Genova (Italy); Martí Duocastella, Istituto Italiano di Tecnologia (Italy), Univ. de Barcelona (Spain)
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Light-sheet microscopes with an extended depth of field (EDOF) offer a simple but powerful route toward fast volumetric imaging. However, methods for EDOF typically result in a loss of signal-to-noise ratio. Here, we propose a parallelization strategy as a simple solution. By illuminating multiple acoustically generated light sheets at different axial positions within the EDOF, and following an encoding sequence, information from several in-focus planes can be simultaneously retrieved. After applying a decoding algorithm, volumetric images are reconstructed with enhanced signal and level of detail. Our strategy paves the way for exploiting the full speed capabilities of EDOF light-sheet systems.
Session 2: High-speed Multiphoton Imaging I
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Author(s): Kaspar Podgorski, Howard Hughes Medical Institute (United States)
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High-frame-rate two-photon imaging is becoming increasingly important in neuroscience for recording fast voltage and neurotransmitter signals from many sources at a time. I will summarize developments in this field and my lab’s recent efforts to build a DMD-based random-access two-photon microscope capable of recording thousands of sources at kilohertz framerates, without the need for computational image reconstruction.
11654-18
Author(s): Shi-Wei Chu, National Taiwan Univ. (Taiwan)
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The brain is one of the most important organs in our body, but it is functionally the least understood one. It is composed of millions of neurons, whose interconnection, i.e. connectome, determines its function. Although the interaction of neurons in vitro has been well studied in the past century, no existing tool can capture whole-brain emergent properties at single neuron or even synapse resolution. To understand functional connectome, an imaging system that can cover a whole brain in vivo with spatial resolution of micrometers (neuron) to nanometers (synapse) as well as temporal resolution in sub-seconds (calcium) to milliseconds (action potential) is highly desirable. In this invited talk, we introduce our recent efforts to improve optical microscopy in terms of speed, depth, and spatial resolution, toward the goal of understanding the brain of Drosophila, which offers a small brain with sophisticated functions and genetic control capabilities.
11654-19
Author(s): Guanghan Meng, Jiang Lan Fan, Qinrong Zhang, Univ. of California, Berkeley (United States); Kevin K. Tsia, The Univ. of Hong Kong (Hong Kong, China); Na Ji, Univ. of California, Berkeley (United States)
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Uncovering the structure and function of cortical vascular network down to the level of capillaries can provide useful insights on brain physiology and pathology. Ideally, the probing method should allow concurrent observation of vascular morphology and hemodynamics, with sufficient spatiotemporal resolution to resolve individual capillaries and track blood cell motion in the scattering mammalian brain. By employing an all-optical scanning two-photon fluorescence microscope, we realized kilohertz full-frame recording of cortical blood vessels beyond 700 µm deep in the mouse brain and measured blood flow speeds up to 15 mm/s
11654-20
Author(s): David Knez, Adam Hanninen, Richard Prince, Dmitry A. Fishman, Eric Potma, Univ. of California, Irvine (United States)
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Nondegenerate two-photon absorption (NTA) is a method for detecting infrared (IR) radiation with wide bandgap photodetectors. We use NTA to detect mid IR (MIR) light on a Si-based detector with the aid of a near-IR (NIR) pump beam. This enables the use of mature, robust Si technologies to detect IR light at room temperature. We show that NTA facilitates MIR spectroscopy in both a single pixel photodiode and a high-resolution CCD, allowing for chemically selective MIR imaging. We demonstrate the utility of NTA by including MIR images and videos of moving targets.
Session 3: High-speed Multiphoton Imaging II
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Author(s): Sebastian Karpf, Nina Gloeckner, Alessandra Henkel, Valerie Lutz, Univ. zu Lübeck (Germany); Bahram Jalali, Univ. of California, Los Angeles (United States); Robert A. Huber, Univ. zu Lübeck (Germany)
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Two-Photon microscopy and fluorescence lifetime microscopy with 4kHz frame-rate is achieved by a spectro-temporal encoded imaging technique. It employs a rapidly wavelength-swept light source which is diffracted on a grating for high-speed line-scans at up to 3.3MHz. The laser, a Fourier-domain mode-locked (FDML) laser, is modulated to short pulses (30ps) by an electro-optical modulator (EOM) and amplified to high peak powers by YDFAs for nonlinear excitation (TPEF, SHG). This active pulse modulation allows for encoding the pixel mapping in time, hence the spectro-temporal encoding allows for digital reconstruction and manipulation of the imaging parameters. By synchronising a high-speed detection system based on sensitive hybrid photodetectors also the transient fluorescence decay curve is aquired, allowing for FLIM imaging at up to 4kHz frame-rate. We present imaging flow cytometry (TPEF, SHG, FLIM) of cell morphologies embedded in unfiltered, whole human blood.
11654-22
Author(s): Mihaela Balu, Alexander Fast, Amanda Durkin, Griffin Lentsch, Beckman Laser Institute and Medical Clinic (United States)
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Multiphoton microscopy enables sub-micron resolution, label-free structural and functional imaging of living tissues with contrast from multiple modalities, including second harmonic generation and two-photon excited fluorescence. We developed a fast, large area multiphoton exoscope (FLAME) portable system with enhanced label-free molecular contrast for macroscopic imaging of human skin with microscopic resolution. It combines optical and mechanical scanning mechanisms with deep learning image restoration to produce 3D sub-cellular resolution images that encompass sub-mm2 to cm2 scale areas of tissue within minutes. We demonstrate the performance and utility of the instrument by fast ex vivo and in vivo imaging of human skin.
11654-23
Author(s): Lisako Kameyama, Sigekazu Takizawa, The Univ. of Tokyo (Japan); Kotaro Hiramatsu, The Univ. of Tokyo (Japan), Japan Science and Technology Agency, PRESTO (Japan); Keisuke Goda, The Univ. of Tokyo (Japan), Univ. California, Los Angeles (United States), Wuhan Univ. (China)
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We introduce a novel method for fast, sensitive dual-comb coherent anti-Stokes Raman (DC-CARS) spectroscopy with a “quasi”-dual-comb laser that achieves an energy efficiency of nearly 100%. Specifically, one of the repetition rates of the two lasers is rapidly modulated by controlling the pump intensity of a Ti:Sapphire laser, while the group delay is monitored with two-color interferometry for the calibration of the time-domain CARS interferogram. With this method, we achieve a record high spectral acquisition rate of 100,000 spectra/s with even higher sensitivity than conventional DC-CARS spectroscopy. Our method holds promise for high-throughput screening, flow cytometry, and live-cell imaging.
11654-24
Author(s): Vincent Maioli, Antoine Boniface, Pierre Mahou, Júlia Ferrer Ortas, Lamiae Abdeladim, Emmanuel Beaurepaire, Willy Supatto, Ecole Polytechnique (France)
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Light-sheet illumination enables major increase in multiphoton imaging speed for in vivo studies. However, photoperturbation in multiphoton light-sheet microscopy remains poorly investigated. We show here that the heart beat rate of zebrafish embryos is a sensitive probe of linear and nonlinear photoperturbations. By analyzing its behavior with respect to laser power, pulse frequency and wavelength, we derive guidelines to balance signal and photoperturbation. We then demonstrate one order-of-magnitude signal enhancement over previous implementations by optimizing the laser pulse frequency. These results open new opportunities for fast in vivo imaging.
11654-25
Author(s): Chien-Sheng Wang, Yu-Hsuan Tsai, Po-Yuan Wang, National Taiwan Univ. (Taiwan); Chih-Wei Liu, Chia-Fu Chou, Academia Sinica (Taiwan); Vijay Raj Singh, Peter T. C. So, Massachusetts Institute of Technology (United States); Shi-Wei Chu, National Taiwan Univ. (Taiwan)
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Here we achieved record-high >500 volumes/second two-photon imaging by improving lateral and axial scanning speed via 32-channel multifocal excitation/detection, and a tunable acoustic gradient-index lens, respectively. We developed a deconvolution process to reduce scattering-induced crosstalk in multifocal detection scheme, thus enabling whole brain imaging of Drosophila with millisecond and micrometer spatiotemporal resolution. Potential applications toward brain science include studying millisecond dynamics in a neuronal network, and resolving 3D microfluidics in blood vessels.
Session 4: Emerging Technologies I
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Author(s): Edward S. Boyden, MIT (United States)
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Understanding and repairing complex biological systems, such as the brain, requires new technologies that enable such systems to be observed and controlled with great precision, across extended spatial and temporal scales. We are discovering new molecular principles that are leading to such technologies. For example, we recently discovered that it was possible to physically magnify biological specimens manyfold, in an even way, by embedding them in dense swellable polymers, mechanically homogenizing the specimens, and then adding water to isotropically swell the specimens. In this method, which we call expansion microscopy (ExM), we enable scalable, inexpensive diffraction-limited microscopes to do large-volume nanoscopy, in a multiplexed fashion – important, for example, for brain mapping. As another example, we discovered that microbial opsins, genetically expressed in neurons, could enable their electrical activities to be precisely driven or silenced in response to millisecond timescale pulses of light. These tools, called optogenetic tools, are enabling causal assessment of the contribution of defined neurons to behaviors and pathologies in a wide variety of basic science settings. Finally, we have developed new methods of directed evolution, and discovered mutant forms of optogenetic tools that enable precision fluorescent imaging of the high-speed voltage of neurons in the living brain. We share all these tools freely, and aim to integrate the use of these tools so as to lead to comprehensive understandings of neural circuits.
11654-8
Author(s): Maria Loidolt-Krüger, Fabian Jolmes, Matthias Patting, Michael Wahl, Evangelos Sismakis, André Devaux, Uwe Ortmann, Felix Koberling, Rainer Erdmann, PicoQuant GmbH (Germany)
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Fluorescence Lifetime Imaging (FLIM) is an essential tool in Life Sciences, but up to now users had to chose between high timing precision or fast data acquisition when using Time-Correlated Single Photon Counting (TCSPC) electronics. Our approach, named rapidFLIMHiRes, allows recording several FLIM images per second with a temporal resolution of 10 ps. The method combines advances in fast scanning, hybrid photomultiplier detectors, TCSPC modules, and correction algorithms to reduce decay curve distortions. Thus fast processes can be observed with the high optical and temporal resolution achievable in confocal microscopy at a rate of several frames per second.
11654-9
Author(s): Yingming Lai, Institut National de la Recherche Scientifique (Canada); Yujia Xue, Boston Univ. (United States); Christian-Yves Côté, Axis Photonique Inc. (Canada); Xianglei Liu, Antoine Laramée, Institut National de la Recherche Scientifique (Canada); Nicolas Jaouen, Synchrotron SOLEIL (France); François Légaré, Institut National de la Recherche Scientifique (Canada); Lei Tian, Boston Univ. (United States); Jinyang Liang, Institut National de la Recherche Scientifique (Canada)
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Compressed ultrafast photography (CUP) is a novel single-shot ultrafast imaging approach that captures an entire transient event with a single exposure. Despite recent developments, CUP has been experimentally demonstrated only in visible and near-infrared spectral ranges. The requirement to tilt a digital mirror device (DMD) in the system and the limited number of controllable parameters in the reconstruction algorithm also hinder CUP’s performance. In this work, we extend CUP to the ultraviolet (UV) spectral range and overcome the previous restrictions caused by applying the tilted DMD. Meanwhile, CUP’s reconstruction quality was enhanced by implementing the plug-and-play (PnP) alternating direction method of multipliers (ADMM) algorithm. The system’s ability of ultrafast imaging was demonstrated by recording the spatially modulated UV pulses. We envision that our system will open up many new possibilities in the observation of transient UV phenomena.
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Author(s): Jongchan Park, Xiaohua Feng, Univ. of California, Los Angeles (United States); Rongguang Liang, The Univ. of Arizona (United States); Liang Gao, Univ. of California, Los Angeles (United States)
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Multidimensional imaging systems, such as hyperspectral and ultrafast imagers, capture optical information beyond 2D spatial intensity distribution by mapping high-dimensional light datacube voxels onto a 2D image sensor. However, the range of applications of the multidimensional imaging system is often restricted by its static optical architecture and measurement scheme. To overcome this limitation, we introduce active optical mapping for multidimensional photography. An active optical mapper, a high-resolution spatial light modulator, map the incident light datacube voxels onto sensor pixels in an arbitrary and programmed manner, thereby maximizing the measurement flexibility. We demonstrate our approach in hyperspectral and high-speed imaging.
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Author(s): Xianglei Liu, Jingdan Liu, Cheng D. Jiang, Fiorenzo Vetrone, Jinyang Liang, Institut National de la Recherche Scientifique (Canada)
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Single-shot real-time ultra-high-speed imaging is of significance in capturing transient phenomena. Existing techniques fall short in possessing satisfying specifications in the imaging speed, sequence depth, and pixel count. To overcome these limitations, we have developed compressed optical-streaking ultra-high-speed photography (COSUP) that records a scene (x, y, t) by applying the operations of spatial encoding, temporal shearing, and spatiotemporal integrating. The COSUP system possesses an imaging speed of 1.5 million frames per second (fps), a sequence depth of 500 frames, and a pixel count of 0.5 megapixels per frame. COSUP is demonstrated by imaging single laser pulses illuminating through transmissive targets and by tracking a fast-moving object. We envision COSUP to be applied in widespread applications in biomedicine and materials science.
Session 5: High-Speed Super-Resolution Imaging
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Author(s): Zhaowei Liu, Univ. of California, San Diego (United States)
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Structured illumination microscope enables high temporal resolution wide field-of-view super-resolution imaging but typically provides only two-fold resolution improvement over the diffraction limit. We present speckle metamaterial-assisted illumination nanoscopy (MAIN) which brings the resolution down to 40 nm and beyond. A hyperbolic metamaterial is implemented as a substrate to generate subwavelength illumination patterns in the near field of the metamaterial. Fluorescent objects located on the metamaterial are illuminated by such high spatial-frequency near field illuminations and are reconstructed by a Blind-SIM algorithm. Speckle-MAIN represents a new route for super-resolution, which may lead to important applications in bio-imaging and surface characterization.
11654-31
Author(s): Ilaria Testa, Science for Life Lab. (Sweden)
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The formidable ability of fluorescent nanoscopy to image features closer than half the wavelength of light often comes at the expense of time and increased dose of energy for recording. RESOLFT is an optical nanoscopy technique that requires only a fraction of the light of for instance STED microscopy, since the fluorescence is inhibited by reversible switching neighbouring molecules into long-lived dark states. By using reversible switchable fluorescent proteins and a novel light pattern we developed a new parallelized imaging system capable of rapid imaging (1 Hz) with super resolution in 3D (< 70 nm). The new interference pattern is obtained by incoherent super imposition of three beam pairs. We validate the system by recording 3D stacks across time of moving organelles and cytoskeleton in living human cells with spatial details < 70 nm in all the three dimensions.
11654-32
Author(s): Xiaofei Han, Lab. of High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health (United States), Tsinghua Univ. (China); Yijun Su, Ryan Christensen, Yicong Wu, Hari Shroff, Lab. of High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering (United States)
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The modern fluorescence microscope would ideally offer high spatial resolution in all dimensions, high speed and minimal photodamage to specimens. We developed a triple-view line confocal system that improves spatial resolution in all three dimensions (to ~270 nm x 250 nm x 335 nm or ~185 nm x 170 nm x 245 nm with structured illumination) in scattering samples tens of microns thick. To speed up acquisition and reduce photobleaching, deep learning is applied to denoise and enhance resolution when only using data acquired from one view.
11654-33
Author(s): Ziqi Zhang, Queenie T. K. Lai, Kelvin C. M. Lee, Kenneth K. Y. Wong, Kevin K. Tsia, The Univ. of Hong Kong (Hong Kong, China)
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We report a large-scale light-scattering single-cell characterization enabled by a high-throughput quantitative phase imaging platform (multi-ATOM) (10,000 cells/sec). By virtue of its subcellular resolution, multi-ATOM accesses the light-scattering information from individual cells via Fourier Transform light scattering (FTLS) analysis. Specifically, we applied FTLS analysis on multi-ATOM images to explore the statistical characteristics of single-cell fractal dimension (FD). We demonstrated that FD can be harnessed as an effective label-free phenotype that is indicative of cell types and states. FD can identify the heterogeneity among leukemia/lung cancer cell types and trace the different phases in cell cycle progression.
Session 6: Imaging Flow Cytometry
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Author(s): Adela Ben-Yakar, The Univ. of Texas at Austin (United States)
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I will present a new fluorescence imaging method called LEAD (line excitation array detection) microscopy, capable of providing 0.8 million frames per second. This method performs 0.8 MHz line-scanning of excitation laser beam using a chirped signal-driven longitudinal acousto-optic deflector to create a virtual light-sheet, and images the field-of-view with a linear photomultiplier tube array to generate a 66×14 pixel frame each scan cycle. I will present an implementation of the LEAD microscopy as a blur-free 3D flow cytometer for Caenorhabditis elegans moving at 1 m/s with 3.5-micron resolution and signal-to-background ratios >200.
11654-35
Author(s): Yoav Shechtman, Elias Nehme, Boris Ferdman, Omer Adir, Racheli Gordon-Soffer, Reut Orange, Tal Naor, Yael Shalev-Ezra, Sarah Goldberg, Onit Alalouf, Technion-Israel Institute of Technology (Israel); Daniel Freedman, Google Haifa (Israel); Avi Schroeder, Tomer Michaeli, Lucien E. Weiss, Technion-Israel Institute of Technology (Israel)
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Attaining three-dimensional data at high throughput is a grand challenge in microscopy. I will discuss two recent contributions to 3D microscopy utilizing point-spread-function (PSF) engineering: 1.a new method that extends the capabilities of flow-based imaging to 3D co-localization microscopy, and 2. efficient and fast localization of dense molecules in volumetric single-molecule-localization microscopy (SMLM) using deep learning, termed DeepSTORM3D. Together, these methods enable the study of sub-cellular biological processes at unprecedented timescales and throughputs.
11654-36
Author(s): Yuqi Zhou, The Univ. of Tokyo (Japan); Atsushi Yasumoto, The Univ. of Tokyo (Japan), Hokkaido Univ. Hospital (Japan); Masako Nishikawa, Yuya Nobori, Ting-Hui Xiao, Yutaka Yatomi, The Univ. of Tokyo (Japan); Masaki Anraku, The Univ. of Tokyo (Japan), Tokyo Metropolitan Geriatric Hospital and Institute of Gerontology (Japan); Keisuke Goda, The Univ. of Tokyo (Japan), Institute of Technological Sciences, Wuhan Univ. (China), Univ. of California, Los Angeles (United States)
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In COVID-19 therapy with artificial lungs such as extracorporeal membrane oxygenation (ECMO) machines, platelets in the extracorporeal circulation are often activated by their contact with the artificial materials, leading to the formation of blood clots followed by serious complications such as stroke and heart attack. However, anticoagulation and antithrombotic management is challenging due to the lack of testing tools to evaluate the circulation. Here we demonstrate real-time monitoring of thrombogenesis in the circulation of an ECMO-equipped goat with an intelligent platelet aggregate characterizer (iPAC), which is based on imaging flow cytometry and deep-learning-based analysis of numerous platelet aggregates in blood.
11654-37
Author(s): Dickson Siu, Michelle C. K. Lo, Kelvin C. M. Lee, Kenneth K. Y. Wong, Michael K. Y. Hsin, James C. M. Ho, Kevin K. Tsia, The Univ. of Hong Kong (Hong Kong, China)
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Using a high-throughput imaging flow cytometer (10,000 cells/sec) multi-ATOM, we established a hierarchical biophysical phenotyping approach for label-free single-cell analysis. We demonstrate that the label-free multi-ATOM contrasts can be derived into a set of spatially hierarchical biophysical features that reflect optical density and dry mass density distributions in local and global scales. This phenotypic profile enables us to delineate subtle cellular response of molecularly targeted drug even at an early time point after the drug administration (6 hours). Based on fluorescence image analysis, we further interpreted how these biophysical phenotypes correlate with specific intracellular organelles alteration upon drug treatment.
Session 7: Emerging Technologies II
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Author(s): Kazuhiro Morimoto, Canon Inc. (Japan); Andrei Ardelean, Arin Can Ulku, Ivan Michel M. Antolovic, EPFL (Switzerland); Valentin Kapitany, Alex Turpin, University of Glasgow (United Kingdom); Claudio Bruschini, EPFL (Switzerland); Daniele Faccio, University of Glasgow (United Kingdom); Edoardo Charbon, EPFL (Switzerland)
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We present a 1 megapixel single-photon avalanche diode (SPAD) camera featuring 3.8 ns time gating and 24 kfps frame rate for 1-bit images, fabricated in 180 nm CMOS image sensor technology. The SPAD sensor was used to capture 2D and 3D scenes over 2 m with depth resolution of 5.4 mm and precision better than 7.8 mm (rms). We demonstrate extended dynamic range in dual exposure operation mode and show spatially overlapped multi-object detection in single-photon time-gated time-of-flight experiments. We further demonstrate applications of the megapixel SPAD camera for fluorescence lifetime imaging microscopy (FLIM) and light-in-flight imaging.
11654-13
Author(s): Jun Ando, RIKEN (Japan)
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We report the high-speed tracking of single biomolecules using metallic nanoparticles as contrast agents. We developed an annular illumination total internal reflection dark-field microscope, and achieved angstrom localization precision and microsecond time resolution with 30 nm and 40 nm gold nanoparticles. We also developed a multicolor dark-field microscope with multiple lasers and a spectrophotometer for illumination and imaging optics, and with silver, gold, and silver-gold alloy nanoparticles as optical probes. I will describe studies, performed by these systems, on fast dynamics of bio-molecular motors such as kinesin and dynein, and on rapid diffusional motions of phospholipids in a supported membrane.
11654-14
Author(s): Qi Cui, Jongchan Park, Univ. of California, Los Angeles (United States); R. Theodore Smith, New York Eye and Ear Infirmary of Mount Sinai (United States); Liang Gao, Univ. of California, Los Angeles (United States)
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The light rays captured by an imaging system contain abundant information, such as spatial coordinates, spectrum and time. However, a traditional camera only detects 2D spatial intensity distribution, throwing away much of the information carried by light rays. To overcome this limitation, we present a snapshot hyperspectral light field imaging system using a single camera. By integrating an unfocused light field camera with a snapshot hyperspectral imager, the image mapping spectrometer, we captured a 5D (x,y,u,v,λ) (x,y, spatial coordinates; u,v, emittance angles; λ, wavelength) datacube in a single camera exposure. We demonstrated the snapshot advantage of our system by imaging the spectral volumetric scenes in real time.
11654-15
Author(s): Meng Cui, Purdue Univ. (United States)
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Towards high-throughput high-quality phosphorescence imaging, we have developed a line scanning mechanical streak camera. Compared to the conventional point scanning based time domain detection, the line scanning mechanical streak camera allows parallel recording from thousands of pixels at high signal to noise ratio. Using a confocal line scanning configuration, we imaged phosphorescence samples with lifetime ranging from tens of nanoseconds to hundreds of microseconds, which demonstrated the simplicity, flexibility, SNR and throughput advantages of the line scanning mechanical streak camera design.
11654-16
Author(s): Allen Kiester, Air Force Research Lab. (United States); Zachary Coker, SAIC (United States); Bennett Ibey, Joel Bixler, Air Force Research Lab. (United States)
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A high performance strobe laser microscopy system has been developed for use in conjunction with FlouVolt™ voltage sensitive dye for imaging variations in cell membrane potential during the application of electric pulses. The system allows flexible variation of trigger timing in order to image any time point of a cell-electric pulse interaction. The system has minimum time resolution of 6 ns and a fixed image exposure time of nominally 6 ns. Electric pulses can be arbitrary in form and phase, with a maximum frequency of 5 MHz, limited by the dye, and maximum intensity of 8 kV/cm.
Session 8: Deep Learning
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Author(s): Anna Kreshuk, European Molecular Biology Lab. (Germany)
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Light field microscopy is a powerful tool for fast volumetric image acquisition in biology which requires a computationally demanding and artefact-prone image reconstruction process. I will present a novel framework consisting of a hybrid light-field light-sheet microscope and deep learning-based volume reconstruction, where single light-sheet acquisitions continuously serve as training data and validation for the convolutional neural network reconstructing the LFM volume. Our framework produces video-rate reconstructions; their fidelity can be verified on demand and the network can be fine-tuned as necessary.
11654-1
Author(s): Michelle C. K. Lo, Kelvin C. M. Lee, Dickson M. D. Siu, Edmund Y. Lam, Kevin K. Tsia, The Univ. of Hong Kong (Hong Kong, China)
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We report a robust method based on generative deep learning to reconstruct quantitative phase image (QPI). By employing multiplexed asymmetric-detection time-stretch optical microscopy (multi-ATOM), we simultaneously captured multiple intensity image contrasts of the same cell in microfluidic flow, revealing different phase-gradient orientations at high throughput (10,000 cells/sec). Using conditional generative adversarial networks (cGAN), we performed a systematic analysis of how different orientations of the phase-gradient contrasts and their combinations influence the QPI prediction performance, which overall general achieves a high similarity (structural similarity index > 0.91) and low error rate (mean squared error < 0.01).
11654-3
Author(s): Dario Polli, Politecnico di Milano (Italy); Alessandro Giuseppi, Sapienza Univ. di Roma (Italy); Federico Vernuccio, Alejandro De la Cadena, Giulio Cerullo, Carlo M. Valensise, Politecnico di Milano (Italy)
Digital Forum: On-demand starting 6 March
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We present a novel approach to remove the unwanted non-resonant background from Broadband Coherent Anti-Stokes Raman Scattering (B-CARS) spectra, based on deep learning. The unsupervised model is built as a convolutional neural network with seven hidden layers. After training on synthetic data, our model was able to process experimental B-CARS spectra and correctly retrieve all the relevant vibrational peaks. The retrieval time is 100 microseconds per spectrum, faster than the time required to record it. We expect that this model will significantly simplify and speed-up the analysis of B-CARS spectra, allowing real-time retrieval of the vibrational features.
11654-4
Author(s): Kangning Zhang, Junjie Hu, Weijian Yang, Univ. of California, Davis (United States)
Digital Forum: On-demand starting 6 March
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We present a new deep compressed imaging modality by scanning a learned illumination pattern on the sample and detecting the signal with a single-pixel detector. This new imaging modality allows a compressed sampling of the object, and thus a high imaging speed. The object is reconstructed through a deep neural network inspired by compressed sensing algorithm. We optimize the illumination pattern and the image reconstruction network by training an end-to-end auto-encoder framework. Comparing with the conventional single-pixel camera and point-scanning imaging system, we accomplish a high-speed imaging with a reduced light dosage, while preserving a high imaging quality.
11654-5
Author(s): Shiyi Cheng, Sipei Fu, Yumi Mun Kim, Boston Univ. (United States); Ji Yi, Johns Hopkins Univ. (United States); Lei Tian, Boston Univ. (United States)
Digital Forum: On-demand starting 6 March
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Standard immunofluorescence (IF) staining is labor-intensive, time-consuming and suffers from inflexibility and poor multiplicity. To overcome these limitations, we proposed a deep learning (DL) approach for virtual IF staining with high multiplicity and specificity from label-free reflectance microscopy. Our results show that DL-enabled label-free IF microscopy can predict characteristic subcellular features during different cell cycles and reveal cellular phenotypes with high accuracy.
Session P: Poster Session
11654-38
Author(s): Yanping He, The Chinese Univ. of Hong Kong (Hong Kong, China); Baoliang Ge, Mo Deng, Massachusetts Institute of Technology (United States); Yijin Wang, Md Habibur Rahman, Yi Ping Ho, Liting Duan, The Chinese Univ. of Hong Kong (Hong Kong, China); Zahid Yaqoob, George Barbastathis, Peter T. C. So, Massachusetts Institute of Technology (United States); Renjie Zhou, The Chinese Univ. of Hong Kong (Hong Kong, China)
Digital Forum: On-demand starting 6 March
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Optical diffraction tomography (ODT) has demonstrated its potential for revealing subcellular structures and quantitative compositions in living cells without chemical staining. Recently, we developed a deep-learning based algorithm to reconstruct the 3D refractive index (RI) maps of cells using a single raw interferogram, measured from an angle-multiplexed ODT system. Using this system, we demonstrated a high throughput 3D image cytometry method, in which a microfluidic chip for controlling cell flow is integrated in the ODT system. By flowing the cells in the chip and minimizing the camera exposure time, we can achieve 3D imaging of over 6,000 cells per second.
11654-39
Author(s): Masaya Okada, Sysmex Corp. (Japan); Yasunori Nawa, Advanced Photonics and Biosensing Open Innovation Lab., National Institute of Advanced Industrial Science and Technology (Japan), Osaka Univ. (Japan); Kazuki Bando, Osaka Univ. (Japan); Yuki Shimaoka, Sysmex Corp. (Japan); Satoshi Fujita, Advanced Photonics and Biosensing Open Innovation Lab., National Institute of Advanced Industrial Sc (Japan), Osaka Univ. (Japan); Katsumasa Fujita, Osaka Univ. (Japan), Advanced Photonics and Biosensing Open Innovation Lab. (Japan); Shigeki Iwanaga, Sysmex Corp. (Japan)
Digital Forum: On-demand starting 6 March
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Amino acids and peptides are basic components of proteins and have vital importance in various biological functions and diseases. In this research, we have attempted to detect and distinguish 20 kinds of amino acids and 39 kinds of peptides without any labeling. By using Raman microscopy, more than two thousand Raman spectra were obtained within five minutes from each analyte, at femtomolecular levels. Furthermore, deep learning analyses of the spectra yielded accuracies greater than 96 percent in discriminating between the amino acids and the peptides.
11654-40
Author(s): Naeeme Modir, Maysam Shahedi, James D. Dormer, The Univ. of Texas at Dallas (United States); Baowei Fei, The Univ. of Texas at Dallas (United States), The Univ. of Texas Southwestern Medical Ctr. at Dallas (United States)
Digital Forum: On-demand starting 6 March
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We designed a mobile, real-time LED-based endoscopic imaging system for the detection of various diseases. The endoscope uses multiple wavelengths within UV, visible, and IR light spectra generated by a micro-LED array. We capture images with a small monochrome camera. A wireless transceiver communicates with a workstation for data delivery and further processing. To investigate the properties and limitations of our high-speed spectral imaging approach, we designed a prototype system. We conducted some experiments to measure the optimal LED forward voltages and lighting duration.
11654-41
Author(s): Tereza Schönfeldová, Sylvie Roke, Ecole Polytechnique Fédérale de Lausanne (Switzerland)
Digital Forum: On-demand starting 6 March
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Dimethylsulfoxide (DMSO) and its mixtures with water are widely used cosolvents and cryoprotectant agents with interesting properties. For example, a DMSO-water mixture with a molar ratio of 1:2 shows a decreased melting point at its eutectic temperature of -75 °C. It is known that DMSO can accept up to two hydrogen bonds and water can be either a donor or an acceptor. However, the mechanism of making and breaking hydrogen-bonds in DMSO-water mixtures is not understood. Here, we used high-throughput femtosecond angle-resolved elastic second harmonic scattering to probe the intermolecular interactions in both pure liquids and their different mixtures.
11654-42
Author(s): Aleksandr S. Grishkanich, LLC Photonics Systems (Russian Federation), LLC Shvabe (Russian Federation); Aleksandr Zhevlakov, S. I. Vavilov State Optical Institute (Russian Federation); Egor Mikharev, Saint Petersburg Electrotechnical University "LETI" (Russian Federation); Andrey Lunev, Saint Petersburg Electrotechnical University (Russian Federation); Boris Karas, Viktoria Pozdniakova, ITMO Univ. (Russian Federation)
Digital Forum: On-demand starting 6 March
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A spectrometer has been developed for high-speed non-invasive diagnostics of diseases of the digestive system using drugs low-enriched in carbon-13 at concentrations of 20%. High-resolution isotope spectrometry system for measuring the concentration of C12 / C13 in non-invasive patient exhalation in real time, which improves the speed and accuracy of measurements. The principle of high-resolution Raman spectroscopy at a wavelength of 532 nm with an integration time of 10 ms is used. Demonstrated analysis of CO2 13C and CO2 12C isotopes with a selectivity of at least 0.1δ ‰ 13C during a test of 10 ms without thermal stabilization systems. The spectrometer is equipped with a software solution for medical decisions.
Front Matter: Volume 11654
Conference Chair
The Univ. of Hong Kong (Hong Kong, China)
Conference Chair
The Univ. of Tokyo (Japan)
Program Committee
Cornell Univ. (United States)
Program Committee
Tsinghua Univ. (China)
Program Committee
National Taiwan Univ. (Taiwan)
Program Committee
Meng Cui
Purdue Univ. (United States)
Program Committee
Qionghai Dai
Tsinghua Univ. (China)
Program Committee
Martí Duocastella
Istituto Italiano di Tecnologia (Italy)
Program Committee
Johns Hopkins Univ. (United States)
Program Committee
Osaka Univ. (Japan)
Program Committee
Univ. of Illinois at Urbana-Champaign (United States)
Program Committee
Citizen Watch Co., Ltd. (Japan)
Program Committee
New Mexico State Univ. (United States)
Program Committee
Bo Huang
Univ. of California, San Francisco (United States)
Program Committee
Univ. of California, Los Angeles (United States)
Program Committee
Pohang Univ. of Science and Technology (Korea, Republic of)
Program Committee
Optores GmbH (Germany)
Program Committee
The Univ. of Hong Kong (Hong Kong, China)
Program Committee
Wuhan Univ. (China)
Program Committee
Univ. of Macau (Macao, China)
Program Committee
Univ. of California, San Diego (United States)
Program Committee
Hokkaido Univ. (Japan)
Program Committee
Columbia Univ. (United States)
Program Committee
CYBO, Inc. (Japan)
Program Committee
Yasushi Okada
RIKEN Quantitative Biology Ctr. (Japan)
Program Committee
KAIST (Korea, Republic of)
Program Committee
Univ. of Kent (United Kingdom)
Program Committee
Politecnico di Milano (Italy)
Program Committee
Univ. of California, Irvine (United States)
Program Committee
Shanghai Institute of Optics and Fine Mechanics, Chinese Academy of Sciences (China)
Program Committee
Massachusetts Institute of Technology (United States)
Program Committee
Boston Univ. (United States)
Program Committee
Wellman Ctr. for Photomedicine (United States)
Program Committee
Univ. of California, Berkeley (United States)
Program Committee
Univ. of Kent (United Kingdom)
Program Committee
Caltech (United States)
Program Committee
The Univ. of Hong Kong (Hong Kong, China)
Program Committee
Yicong Wu
National Institutes of Health (United States)
Program Committee
Tokushima Univ. (Japan)
Program Committee
Tomokazu Yoshida
Sysmex Corp. (Japan)
Program Committee
Bar-Ilan Univ. (Israel)
Additional Information
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Post-Deadline Submission Portal