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检索条件"任意字段=Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing IX"

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three-dimensional tomographic imaging and quantitative phase measurement using virtual phase conjugation 27

Three-dimensional tomographic imaging and quantitative phase...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XXVII

作者： Kawashima, Satoshi Okamoto, Atsushi Maeda, Tomohiro Ogawa, Kazuhisa Tomita, Akihisa Osaki, Ryusuke Hayashi, Naoki Hokkaido Univ Grad Sch Informat Sci & Technol Kita Ku Kita14 Nishi9 Sapporo Hokkaido 0600814 Japan FUJIFILM Corp Minato Ku 7-3 Akasaka 9 Chome Tokyo 1070052 Japan

ISBN: (纸本)9781510632547

We have proposed virtual-phase-conjugation-based optical tomography (VPC-OT) using a virtual phase conjugation technology for single-shot and three-dimensional optical tomography. In VPC-OT, a random-spatial-phase-modulated probe beam is irradiated to the sample to be measured, and the complex amplitude of the signal composed of a superimposition of light reflected from each layer of the sample is measured. A three-dimensional tomogram of intensity and phase is obtained by reproducing the measured complex amplitude using a phase conjugate wave in a virtual optical system built in a computer. At this time, by changing the parameters of the virtual optical system, it becomes possible to obtain information of various tomographic planes from the data obtained with a single measurement. In the ideal virtual phase conjugate reproduction process, free space propagation can be assumed;however, in the actual measurement, due to the distortion of the waves and the surroundings of the sample to be measured, a mismatch will occur in modulation and demodulation, and the separation accuracy between different tomographic planes would be degraded. We perform an experiment to clarify the characteristics of VPC-OT in this situation. In this experiment, three-dimensional optical tomography is performed using an etching glass having a periodic structure of 30 mu m as a sample, and the phase distribution is measured quantitatively. Furthermore, by placing a cover glass in front of the object and performing the same measurement, we discuss the characteristics and performance of VPC-OT when there is an optical distortion around the sample to be measured.

关键词： Phase conjugation digital holography tomographic image processing tomography three-dimensional image acquisition wavefront encoding

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A multiplexed confocal FLIM microscope with 4-taps time-gated imager 27

A multiplexed confocal FLIM microscope with 4-taps time-gate...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XXVII

作者： Richards, Morgan Shirakawa, Yuya Badr, Fares Kagawa, Keiichiro Kawahito, Shoji Fang, Qiyin McMaster Univ Dept Engn Phys 1280 Main St West Hamilton ON L8S 4L7 Canada Shizuoka Univ Res Inst Elect 3-5-1 Johoku Hamamatsu Shizuoka 4328011 Japan McMaster Univ Sch Biomed Engn 1280 Main St West Hamilton ON L8S 4L7 Canada

ISBN: (纸本)9781510632547

Combined with confocal imaging, Fluorescence lifetime imaging microscopy (FLIM) can achieve 3-dimensional optical sectional capability with sub-nanosecond lifetime information. As confocal FLIM acquires multi-dimensional data 4D (3D space + time), it is inherently slow. Recent developments in lock-in pixel imagers with time gated pixels show such detectors are capable of collecting as many as 8-time gates in a single pixel cycle. We present a multiplexed confocal FLIM microscope, equipped with a 4-taps time-gated lock-in pixel imager. The multiplexing setup allows the use of the sparse array with sub-nanosecond time-gating to achieve high throughput FLIM acquisition.

关键词： Multipoint Confocal Fluorescence FLIM Time-resolved CMOS image sensor lock-in pixel biomedical imaging

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Compact spectrograph for hyperspectral imaging with overlapping fluorescent reporters 27

Compact spectrograph for hyperspectral imaging with overlapp...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XXVII

作者： Hart, Nathan A. Gibbs, Holly C. Yeh, Alvin T. Texas A&M Univ Dept Biol Dept Biomed Engn College Stn TX 77843 USA

ISBN: (纸本)9781510632547

A compact, prismbased spectrograph was designed for 2-photon light sheet microscopy based on broadband, ultrashort optical pulses that corrects for chromatic aberrations and distortion. Gaussian or Bessel beams of broadband optical pulses may be used to create narrow cylindrical, nonlinear excitation volumes from which fluorophores emit characteristic spectra. In practice, a slit aperture is often used to optically section the excitation volume before being imaged onto the camera chip. Optical dispersion of the fluorescence in the transverse direction of the imaging slit allows for hyperspectral image acquisition. Hyperspectral imaging systems may be used to simultaneously image and segment multiple fluorescent reporters in biological tissue. However, current systems have properties which are undesirable for low-light microscopy including chromatic aberrations, distortion, low optical transmission, and large footprint which consumes precious laboratory real estate. Here, we present a novel spectrograph that has sufficient optical transmission, achromaticity, and distortion correction for microscopy of fluorescent reporters spanning the visible spectrum (400 650 nm). Across the spatial dimension of the excitation volume, the spectrograph has uniform separation of the spectral bands while maintaining a compact size and profile. A second order deconvolution algorithm is used to spectrally deconvolve overlapping fluorophores.

关键词： broadband excitation scanning plane microscopy multiplexed microscopy hyperspectral imaging spectral deconvolution

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3D refractive index tomography with quantitative oblique back-illumination microscopy 27

3D refractive index tomography with quantitative oblique bac...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XXVII

作者： Ledwig, Patrick B. Robles, Francisco E. Georgia Inst Technol Atlanta GA 30332 USA Emory Univ Sch Med Atlanta GA 30322 USA

ISBN: (纸本)9781510632547

We have recently developed quantitative oblique back-illumination microscopy (qOBM), which enables full-field quantitative phase imaging (QPI) of objects embedded in a thick scattering medium. This epi-mode technique makes use of multiple scattering as a source of transmissive illumination from within, allowing for rich structural detail based on optical path delay. We now produce quantitative 3D renderings of index of refraction with subcellular detail by computing a 3D transfer function of the entire optical system, including the multiply scattered illumination, and deconvolving it from a vertical stack of phase gradient contrast images. This approach allows truly non-invasive, label-free, tomographic quantitative reconstructions if index of refraction in thick scattering samples including thick tissue samples, at low cost and with simple operation, bringing QPI's unparalleled access to sub-cellular structural detail to previously unavailable domains of investigation.

关键词： back illumination quantitative phase thick sample refractive index multiple scattering differential phase contrast microscopy deconvolution

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Super-speed multiphoton microscopy for mesoscopic volume imaging with ultra-dense sampling beyond Nyquist Limit 27

Super-speed multiphoton microscopy for mesoscopic volume ima...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XXVII

作者： Borah, Bhaskar Jyoti Chi, Han-Hsiung Yen, Chen-Tung Sun, Chi-Kuang Natl Taiwan Univ Dept Elect Engn Taipei 10617 Taiwan Natl Taiwan Univ Grad Inst Photon & Optoelect Taipei 10617 Taiwan Natl Taiwan Univ Dept Life Sci Taipei 10617 Taiwan Natl Taiwan Univ Coll Med Taipei 10051 Taiwan Natl Taiwan Univ Mol Imaging Ctr Taipei 10617 Taiwan Natl Taiwan Univ Grad Inst Biomed Elect & Bioinformat Taipei 10617 Taiwan Acad Sinica Res Ctr Appl Sci Taipei 11529 Taiwan

ISBN: (纸本)9781510632547

We developed a nonlinear optical microscopy (NLOM) system with an extended field of view (FOV) of up -to 1.6 mm x 1.6 mm, employing a high numerical aperture (NA) and low magnification objective lens (20x, NA>0.9) maintaining submicron lateral resolution and <2 mu m axial resolution, making the system a suitable candidate for high resolution imaging. An acquisition speed of 95 M samples per second was implemented with synchronized sampling of 1 voxel per optical pulse from a femtosecond laser source with a 95 MHz repetition rate, with the ability of scanning a 1.6 x 1.6 x 1.6 mm(3) volume, with 12000 x 12000 x 2000 (x 4 channels), i.e., 1.152 Tera-voxels in total, capturing similar to 2.1 Terabyte of 16-bit raw data with 14-bit resolution in <53 minutes at 0.8 mu m Z-steps, and maintaining a Nyquist-exceeded voxel-size, a Nyquist-exceeded volume-scanning speed and a Nyquist-exceeded line-scanning speed of <15 attoliter, >1288 mu m(3)/ms and >12 mm/ms, respectively.

关键词： nonlinear optical microscopy field of view high resolution imaging sampling rate Nyquist-exceeded voxel-size

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Excitation-scan Mirror Array System Advancements to Hyperspectral Imaging Applications 27

Excitation-scan Mirror Array System Advancements to Hyperspe...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XXVII

作者： Browning, Craig M. Mayes, Sam A. Deal, Joshua Gunn-Mayes, Samantha Rich, Thomas C. Leavesley, Silas J. Parker, Marina Univ S Alabama Chem & Biomol Engn Mobile AL 36688 USA Univ S Alabama Pharmacol Mobile AL 36688 USA Univ S Alabama Ctr Lung Biol Mobile AL 36688 USA Univ S Alabama Dept Syst Engn Mobile AL 36688 USA

ISBN: (纸本)9781510632547

Hyperspectral imaging (HSI) technology has been applied in a range of fields for target detection and mixture analysis. While its original applications were in remote sensing, modern uses include agriculture, historical document authentications and medicine. HSI has shown great utility in fluorescence microscopy;however, acquisition speeds have been slow due to light losses associated with spectral filtering. We are currently developing a rapid hyperspectral imaging platform for 5-dimensional imaging (RHIP-5D), a confocal imaging system that will allow users to obtain simultaneous measurements of many fluorescent labels. We have previously reported on optical modeling performance of the system. This previous model investigated geometrical capability of designing a multifaceted mirror imaging system as an initial approach to sample light at many wavelengths. The design utilized light-emitting diodes (LEDs) and a multifaceted mirror array to combine light sources into a liquid light guide (LLG). The computational model was constructed using Monte Carlo optical ray software (TracePro, Lambda Research Corp.). Recent results presented here show transmission has increased up to 9% through parametric optimization of each component. Future work will involve system validation using a prototype engineered based on our optimized model. System requirements will be evaluated to determine if potential design changes are needed to improve the system. We will report on-spectral resolution to demonstrate feasibility of the RHIP-5D as a promising solution for overcoming current HSI acquisition speed and sensitivity limitations.

关键词： Systems Design LED Tissue imaging HSI Spectroscopy microscopy Transmission Spectral Fluorescence Molecules Cells

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Refractive index properties of the retina accessed by multi-wavelength digital holographic microscopy 26

Refractive index properties of the retina accessed by multi-...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XXVI

作者： Barroso, Alvaro Ketelhut, Steffi Heiduschka, Peter Nettels-Hackert, Gerburg Schnekenburger, Juergen Kemper, Bjoern Univ Munster Med Fac Biomed Technol Ctr Mendelstr 17 D-48149 Munster Germany Univ Munster Dept Ophtalmol Med Ctr Domagkstr 15 D-48149 Munster Germany

ISBN: (纸本)9781510624092

The refractive index (RI) of the retina and its dispersion are essential parameters in ophthalmologic imaging. However, the spatial RI distribution in retinal tissue is difficult to access. We explored the capabilities of multispectral quantitative phase imaging (QPI) with digital holographic microscopy (DHM) for label-free refractive index characterization of dissected murine retina. The retrieved tissue refractive indices are in agreement with previously reported values for living cells and dissected tissues. Moreover, the detected spatial refractive index distributions correlate with results from complementary conducted OCT investigations. In summary, multispectral DHM is a promising tool for label-free characterization of optical retina properties.

关键词： digital holographic microscopy quantitative phase imaging multi-wavelength hyperspectral imaging retina refractive index

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Enhanced spectral lightfield fusion microscopy via deep computational optics for whole-slide pathology 26

Enhanced spectral lightfield fusion microscopy via deep comp...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XXVI

作者： Kazemzadeh, Farnoud Li-Chang, Hector Wong, Alexander Univ Waterloo Waterloo ON Canada Waterloo Artificial Intelligence Inst Waterloo ON Canada Royal Victoria Reg Hlth Ctr Barrie ON Canada

ISBN: (纸本)9781510624092

Lensfree on-chip microscopy, which harnesses holography principles to capture interferometric light-field encodings without the need of lenses, is an emerging microscopy modality with widespread interest given the large field-of-view (FOV) compared to lens-based microscopy systems. In particular, there is a growing interest on the development of high-quality lensfree on-chip color microscopy. In this study, we propose a multi-laser spectral lightfield fusion microscopy using deep computational optics for achieving lensfree on-chip color microscopy. We will demonstrate that leveraging deep computational optics can enable imaging resolution beyond the diffraction limit without the use of any complex hardware-based super-resolution techniques, such as aperture scanning. The capabilities of the microscope are examined for whole-slide pathology. The superior imaging resolution of the instrument is demonstrated by imaging of a series of biological specimens demonstrating the true color imaging capability of the instrument while showcasing the large FOV of the instrument.

关键词： on-chip microscopy deep computational optics light-field fusion

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Temporal focusing-based multiphoton excitation fluorescence images with background noise cancellation via Hilbert-Huang transform 26

Temporal focusing-based multiphoton excitation fluorescence ...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XXVI

作者： Hu, Yvonne Yuling Luo, Yuan-Rong Lin, Chun-Yu Chang, Chia-Yuan Chen, Shean-Jen Natl Cheng Kung Univ Dept Photon Tainan Taiwan Natl Cheng Kung Univ Dept Engn Sci Tainan Taiwan Natl Cheng Kung Univ Dept Mech Engn Tainan Taiwan Natl Chiao Tung Univ Inst Imaging & Biomed Photon Coll Photon Tainan Taiwan

ISBN: (纸本)9781510624092

Temporal focusing multiphoton excitation microscopy has wide field-of-view and optical sectioning. By using digital micromirror device, it provides patterned illumination. However, without filling the back aperture of objective lens, the axial confinement is limited to micron-meters, leading the out-of-focus fluorophores excited and image blurred. In this study, Hilbert-Huang transform is proposed to reduce the background noise. The empirical mode decomposition is first applied to disassemble the image into intrinsic mode functions and then reconstruct by Hilbert transform after diminishing background residues. The axial confinement can be enhanced from 2.79 mu m to 0.73 mu m with structure frequency in 1.06 mu m(-1).

关键词： Temporal focusing Digital micromirror device Structured illumination microscopy Hilbert-Huang transform

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Wave-optical calibration of a light-field fluorescence microscopy 26

Wave-optical calibration of a light-field fluorescence micro...

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Conference on three-dimensional and multidimensional microscopy - image acquisition and processing XXVI

作者： Pribosek, J. Steinbrener, J. Baumgart, M. Bereczki, T. Harms, K. Tortschanoff, A. Kenda, A. Carinthian Tech Res Europastr 12 A-9524 Villach Austria Mol Devicess Lagerhausstr 45 A-5071 Wals Austria

ISBN: (纸本)9781510624092

In light-field microscopy, a single point emitter gives rise to a complex diffraction pattern, which varies with the position of the emitter in object space. In order to use deconvolution-based wave-optical reconstruction schemes for light-field imaging systems, established methods rely on theoretical estimation of such diffraction patterns. In this paper we propose a novel method for direct experimental estimation of the light-field point spread function. Our approach relies on a modified reversed micro-Hartmann test to acquire a composite light-field point spread function of several thousand point emitters in the object plane simultaneously. By using fiducial markers and a custom image processing algorithm we separate the contributions of individual point emitters directly in raw light-field images and allow the construction of the forward imaging process without any prior assumption about the optical system required. The constructed forward imaging model can finally be applied in the 3D-deconvolution based wave-optical reconstruction scheme.

关键词： light-field microscopy light-field point spread function wave-optical calibration fluorescence imaging

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