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Lensfree on-chip high-resolution imaging using two-way lighting, and its limitations 13

Lensfree on-chip high-resolution imaging using two-way light...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXIII

作者： Adachi, Yasuhiko Tamaki, Tokuhiko Motomura, Hideto Kato, Yoshihisa Panasonic Corp Adv Res Div Osaka Japan Panasonic Corp Automot & Ind Syst Co Div Engn Kyoto Japan

ISBN: (纸本)9781628419474

A high-magnification image of a biological sample can generally be obtained by an optical microscope with an objective lens, moving the image sensor with a sub-pixel shift and the subsequent image processing for super-resolution. However, to obtain a high-resolution image, a large number of images will be required for the super-resolution, and thus it is difficult to achieve real-time operation, and the field-of-view (FOV) is not sufficiently wide. The currently proposed digital holography technique places a sample on the image sensor and captures the interference fringe (hologram) to reconstruct a 3D high-resolution image in a computer. This technique ensures the features of a wide FOV, whereas the high resolution obtained by image processing cannot ensure real-time operation, because it requires recursive calculations of light propagation and adequate computer resources. To realize wide FOV and the real-time operation at the same time, we have developed a new technique: Lensfree on-chip high-resolution imaging using two-way lighting. High-resolution image is immediately obtained by image processing of the low-resolution images of the samples. This makes it possible to ensure a wide FOV, a deep depth of focus without the need for focus adjustment, and a continuously expanding operation. We also discuss the limitations of the high resolution.

关键词： Lensfree microscopy On-chip microscopy Two-way lighting

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Time-resolved wide-field optically sectioned fluorescence microscopy

Time-resolved wide-field optically sectioned fluorescence mi...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XX

作者： Dupuis, Guillaume Benabdallah, Nadia Chopinaud, Aurelien Mayet, Celine Leveque-Fort, Sandrine Univ Paris 11 CPBM Federat LUMAT FR 2764 Orsay France CNRS Orsay France Univ Paris 11 Inst Sci Mol Orsay UMR 8214 Orsay France

ISBN: (纸本)9780819493583

We present the implementation of a fast wide-field optical sectioning technique called HiLo microscopy on a fluorescence lifetime imaging microscope. HiLo microscopy is based on the fusion of two images, one with structured illumination and another with uniform illumination. Optically sectioned images are then digitally generated thanks to a fusion algorithm. HiLo images are comparable in quality with confocal images but they can be acquired faster over larger fields of view. We obtain 4-D imaging by combining HiLo optical sectioning, time-gated detection, and z-displacement. We characterize the performances of this set-up in terms of 3-D spatial resolution and time-resolved capabilities in both fixed- and live-cell imaging modes.

关键词： fluorescence microscopy fluorescence lifetime imaging microscopy (FLIM) structured illumination microscopy (HiLo) wide-field optical sectioning

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Two-dimensional compressive sensing in spectral domain optical coherence tomography 22

Two-dimensional compressive sensing in spectral domain optic...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXII

作者： Xu, Daguang Huang, Yong Kang, Jin U. Johns Hopkins Univ Dept Elect & Comp Engn Baltimore MD 21218 USA

ISBN: (纸本)9781628414202

In this paper, we proposed a novel compressive sensing (CS) method in spectral domain optical coherence tomography (SD OCT), which reconstructs B-scan image using a subset of the spectral data that is under-sampled in both axial and lateral dimensions. Thus a fraction of the A-scans for a B-scan are acquired;the spectral data of each acquired A-scan is under-sampled. Compared with the previous studies, our method further reduces the overall size of the spectral measurements. Experimental results show that our approach can obtain high quality B-scan image using 25% spectral data, which takes 50% number of A-scans and acquires 50% spectral data for each selected A-scan.

关键词： Compressive sensing optical coherence tomography reconstruction algorithm image processing

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Transmission confocal microscopy: making it a reality

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conference on 3-dimensional and multidimensional microscopy - image acquisition and processing V

作者： Arnison, MR Fekete, PW Serrano, M Nguyen, PTA Romagnoli, R Guan, L O'Byrne, JW Cogswell, CJ Univ Sydney Sch Phys Dept Phys Opt Sydney NSW 2006 Australia

ISBN: (纸本)0819427004

Imaging thick specimens in 3D transmission confocal modes presents two key problems. The first problem is variable aberrations introduced by changes in refractive index. The second problem is revealed when visualising acquired data, where thick 3D datasets are difficult to interpret. In this paper we present our emerging solutions to these problems. Aberrations can be classified as simple tip-tilt deflection of the beam, or more complicated higher order aberrations. We discuss our results which demonstrate successful on-the-fly detection and correction for tip-tilt For detecting higher order aberrations, we have chosen to investigate the wavefront curvature sensing technique. The second problem of rendering thick 3D datasets can be solved by extracting features of interest from the background. Simple intensity threshholding is not sufficient for complex biological specimens, and image processing in only two dimensions neglects any three dimensional structure. Use of Kohonen's self-organising map neural network in 3D results in clear segmentation of features for sample chromosome specimens.

关键词： 3D microscopy confocal transmission DIC adaptive optics neural network image processing

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Dynamic phase imaging and processing of moving biological organisms

Dynamic phase imaging and processing of moving biological or...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing Xix

作者： Creath, Katherine Goldstein, Goldie 4D Technol Corp Tucson AZ 85706 USA

ISBN: (纸本)9780819488701

This paper describes recent advances in developing a new, novel interference Linnik microscope system and presents images and data of live biological samples. The specially designed optical system enables instantaneous 4-dimensional video measurements of dynamic motions within and among live cells without the need for contrast agents. "Label-free" measurements of biological objects in reflection using harmless light levels are possible without the need for scanning and vibration isolation. This instrument utilizes a pixelated phase mask enabling simultaneous measurement of multiple interference patterns taking advantage of the polarization properties of light enabling phase image movies in real time at video rates to track dynamic motions and volumetric changes. Optical thickness data are derived from phase images after processing to remove the background surface shape to quantify changes in cell position and volume. Data from a number of different pond organisms will be presented, as will measurements of human breast cancer cells with the addition of various agents that break down the cells. These data highlight examples of the image processing involved and the monitoring of different biological processes.

关键词： phase imaging interference microscopy polarization interferometry cellular imaging cell dynamics optical thickness measurement label-free imaging 4D microscopy

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Bright field segmentation tomography (BFST) for use as surface identification in stereomicroscopy

Bright field segmentation tomography (BFST) for use as surfa...

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conference on 3-dimensional and multidimensional microscopy - image acquisition and processing XI

作者： Thiesse, J Namati, E de Ryk, J Hoffman, E McLennan, G Univ Iowa Iowa City IA 52242 USA

ISBN: (纸本)0819452327

Stereomicroscopy is an important method for use in image acquisition because it provides a 3D image of an object when other microscopic techniques can only provide the image in 2D. One challenge with this type of imaging is determining the top surface of a sample that has otherwise indistinguishable surface and planar characteristics. We have developed a system that creates a form of oblique illumination and in conjunction with image processing, the top surface can be viewed. The use of BFST consists of the Leica MZ16A stereomicroscope with a unique attached lighting source, and is part of the Large image Microscope Array (LIMA) [1]. The lighting source consists of eight light emitting diodes (LED's) that are separated by 45-degree angles. Each LED in this system illuminates with a 20-degree viewing angle once per cycle with a shadow over the rest of the sample. Subsequently, eight segmented images are taken per cycle. After the images are captured they are stacked through image addition to achieve the full field of view, and the surface is then easily identified. image processing techniques, such as skeletonization, can be used for further enhancement and measurement. With the use of BFST, advances can be made in detecting surface features from metals to tissue samples, such as in the analytical assessment of pulmonary emphysema using mean linear intercept.

关键词： stereomicroscopy illumination surface identification image processing

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Comparing methods for 3D microscopy

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conference on 3-dimensional and multidimensional microscopy - image acquisition and processing V

作者： McNally, JG Markham, J Conchello, JA Washington Univ Inst Biomed Comp St Louis MO 63110 USA

ISBN: (纸本)0819427004

In previous studies, we examined three-dimensional images of a latex bead containing a surface layer of fluorescence whose thickness was determined by physical sectioning. Confocal images of the bead from six different microscopes all exhibited a significantly thicker fluorescent shell than actually present. In contrast, deconvolved wide-field images of the bead produced an image with the correct shell thickness. We have now repeated some of these studies using a new latex bead containing a much thinner layer of surface fluorescence. In contrast to earlier studies, confocal images of this bead appear to show an accurate thickness for the fluorescent shell. These particular confocal microscopes (a Biorad 1024 and a Zeiss 510) also yielded essentially aberration free images, which was not the case for some of the earlier microscopes tested. In these recent studies however, deconvolution of the bead appeared less robust than earlier. Of three independent wide-held images of the bead, only one yielded a substantially artifact-free image upon deconvolution.

关键词： 3D microscopy comparison confocal wide held deconvolution

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Phase imaging digital holography for biological microscopy

Phase imaging digital holography for biological microscopy

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conference on 3-dimensional and multidimensional microscopy - image acquisition and processing XI

作者： Kim, MK Parshall, D Univ S Florida Dept Phys Tampa FL 33620 USA

ISBN: (纸本)0819452327

The digital holographic microscopy is being developed, to offer a number of significant advantages and capabilities in phase imaging. The direct availability of both the amplitude and phase information offers a range of versatile processing techniques that can be applied to the complex field data, including the phase imaging which is particularly straightforward in digital holography. One of the techniques we are developing addresses the problem of 2-pi phase discontinuity in the phase image, where most of the conventional phase unwrapping algorithms require subjective intervention for multi-lambda discontinuity. In the present system, we generate two phase maps by digital holography using two different wavelengths. Numerical superposition of the two phase maps results in a new phase map whose effective wavelength is inversely proportional to the difference of the two wavelengths. The axial range can in principle be arbitrarily large compared to the wavelength, while maintaining axial resolution to a fraction of wavelength. We apply the phase imaging digital holography to a number of systems, including the imaging of thin films and biological cells. Preliminary results from these experiments are presented and future development and applications are discussed.

关键词： phase imaging digital holography phase unwrapping microscopy multi-wavelength imaging

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Adaptive compensation of aberrations in ultrafast 3D microscopy using a deformable mirror

Adaptive compensation of aberrations in ultrafast 3D microsc...

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conference on 3 dimensional and multidimensional microscopy - image acquisition processing VII

作者： Sherman, L Albert, O Schmidt, C Vdovin, G Mourou, G Norris, TB Univ Michigan Ctr Ultrafast Opt Sci Ann Arbor MI 48109 USA

ISBN: (纸本)081943535X

3D imaging using a multiphoton scanning confocal microscope is ultimately Limited by aberrations of the system. We describe a system to adaptively compensate the aberrations with a deformable mirror. We have increased the transverse scanning range of the microscope by three with compensation of off-axis aberrations. We have also significantly increased the longitudinal scanning depth with compensation of spherical aberrations from the penetration into the sample. Our correction is based on a genetic algorithm that uses second harmonic or two-photon fluorescence signal excited by femtosecond pulses from the sample as the enhancement parameter. This allows us to globally optimize the wavefront without a wavefront measurement. To improve the speed of the optimization we use Zernike polynomials as the basis for correction. Corrections can be stored in a database for look-up with future samples.

关键词： scanning confocal microscopy aberration correction

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A compact open-top light-sheet microscope for Optofluidic imaging 29

A compact open-top light-sheet microscope for Optofluidic im...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXix

作者： Son, Jeonghwan Mandracchia, Biagio Jia, Shu Georgia Inst Technol Wallace H Coulter Dept Biomed Engn Atlanta GA 30332 USA Emory Univ Atlanta GA 30332 USA

ISBN: (纸本)9781510648043;9781510648036

The fluorescence Optofluidic imaging have recently transformed conventional optical investigations of biological systems by exploiting the synergic combination of recent microscopy and integrated chips technologies. The implementation of light-sheet microscopy on Optofluidic imaging has further pushed the systemic capabilities towards fast and 3D-volumetric imaging. Here, we propose a new compact open-top light-sheet microscope to extend the study of 3D-fluorescence Optofluidic imaging. The proposed portable architecture of the system and customized reconstruction algorithms enable providing high throughput and volumetric reconstruction capability with an isometric single-cell resolution. The experimental results demonstrate the applicability of open-top light-sheet configuration to single-cell imaging in a flow. We anticipate the system to offer promising applications in clinical settings where the portability and 3D-volumetric analysis are required for fast diagnostics.

关键词： Light-sheet microscopy Open-top Light-sheet microscopy Portable microscopy Optofluidic Optofluidic imaging 3D-Volumetric imaging Volumetric imaging High-throughput imaging

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