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Conference on 3-dimensional and multidimensional microscopy - image acquisition and processing X

作者： Genovesio, A Olivo-Marin, JC Inst Pasteur Quantitat Image Anal Grp F-75724 Paris France

ISBN: (纸本)0819447641

The analysis of cell and pathogen movement and motility is a major topic in cell biology for which computerized methods are most needed. This study proposes a method to detect and track multiple moving biological objects in image sequences acquired through fluorescence video microscopy. The method enables the analysis of video microscopy image sequences in order to obtain reliable quantitative data such as number, position, speed and movement phases. The method consists of three stages. A stage of detection is performed through a multi-scale analysis of images using an undecimated wavelet transform. The next stage is the prediction of the state of each detected spot in the next frame using a Kalman filter and an adapted model. Then comes a stage of data association which constructs the tracks and refines the filters. Once all moving objects have been assigned with unique spatio-temporal paths, trajectories are analyzed in terms of different parameters relevant to the motility analysis of biological objects.

关键词： image processing tracking wavelets Kalman filter data associations biological video microscopy fluorescent spots

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Hyper-spectrum scanning laser optical tomography 25

Hyper-spectrum scanning laser optical tomography

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Conference on three-dimensional and multidimensional microscopy: image acquisition and processing XXV

作者： Chen, Lingling Li, Guiye Li, Yingchao Liu, Lina Liu, Ang Hu, Xuejuan Ruan, Shuangchen Shenzhen Univ Coll Optoelect Engn Shenzhen 518060 Peoples R China

ISBN: (数字)9781510614840

ISBN: (纸本)9781510614840;9781510614833

We describe a quantitative fluorescence projection tomography technique which measures the three-dimensional fluorescence spectrum in biomedical samples with size up to several millimeters. This is achieved by acquiring a series of hyperspectral images, by using laser scanning scheme, at different projection angles. We demonstrate that this technique provide a quantitative measure of the fluorescence signal by comparing the spectrum and intensity profile of a fluorescent bead phantom and also demonstrate its application to differentiating the extrinsic label and the autofluorescence in a mouse embryo.

关键词： Fluorescence microcopy three-dimensional microscopy Multi-dimensional imaging Imaging reconstruction techniques Spectrum-resolved imaging

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Automatic quantification of neurite outgrowth by means of image analysis

Automatic quantification of neurite outgrowth by means of im...

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

作者： Van de Wouwer, G Nuydens, R Meert, T Weyn, B Univ Antwerp Visielab Dept Phys B-2020 Antwerp Belgium

ISBN: (纸本)0819452327

A system for quantification of neurite outgrowth in in-vitro experiments is described. The system is developed for routine use in a high-throughput setting and is therefore needs fast, cheap, and robust. It relies on automated digital microscopical imaging of microtiter plates. image analysis is applied to extract features for characterisation of neurite outgrowth. The system is tested in a dose-response experiment on PC12 cells + Taxol. The performance of the system and its ability to measure changes on neuronal morphology is studied.

关键词： automation microscopy image analysis neurite outgrowth

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Use of bessel beams and position-sensitive detectors in scanning photon microscope for improved field depth and contrast

Use of bessel beams and position-sensitive detectors in scan...

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

作者： Khmaladze, Alexander Kim, Myung K. Univ S Florida Dept Phys 4202 E Fowler Ave Tampa FL 33620 USA

ISBN: (纸本)0819461326

The scanning photon microscope technique is a method of microscopic image formation that employs a laser beam focused on a sample, while non-imaging detector receives the scattered light. The scans are achieved by means of a galvanometer based scanning mirror and a motorized micrometer. The system produces images analogous to the scanning electron microscopy with three-dimensional effects of shadowing and reflection. Compared to a conventional wide-field imaging system, the method allows for a greater ease of operation and flexibility, as the image quality is dependent upon the characteristics of the laser beam, rather than imaging optics. The image resolution on the order of a micron is demonstrated. A further gain in terms of resolution and the depth of focus by employment of Bessel rather than Gaussian beams is discussed. Additionally, we used a position-sensitive quadrant photodiode detector to highlight the overall spatial orientation of the imaged surface as well as its roughness. This concept can be useful in many areas, such as coherence imaging and fluorescence.

关键词： scanning microscopy axicon surface profile

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Programmable beam-splitter for confocal laser scanning microscopy

Programmable beam-splitter for confocal laser scanning micro...

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Conference on 3-dimensional and multidimensional microscopy: image acquisition and processing ix

作者： Birk, H Engelhardt, J Storz, R Hartmann, N Bradl, J Ulrich, H Leica Microsyst Heidelberg GMBH D-68165 Mannheim Germany

ISBN: (纸本)0819443603

In confocal microscopy manifold combinations of dyes are used. The entire wavelength range from near-ultraviolet to near-infrared is used to excite these dyes. Their emission is then detected primarily in the visible range. In addition to the number of dyes, the number of possible excitation laser lines is already large and constantly increasing. Due to the almost unlimited number of possible combinations, programmable devices are required. This is true for excitation-modules, beam-splitters and detection modules. Systems using filters with fixed spectral properties can practically fulfill the requirements for only a very small subset of applications. Programmable devices have been realized for excitation using Acousto Optical Tunable Filters (AOTF's). Freely definable spectral detectors are available for a perfect adaptation to the desired emission bands. The missing link for a completely filter-free design is now introduced using an Acousto Optical Beam Splitter (AOBS(TM)).

关键词： multi-spectral fluorescence detection Acousto-Optical Tunable Filter (AOTF) Acousto Optical Beam Splitter (AOBS((TM))) confocal laser scanning

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Multi-wavelength digital holographic tomography based on spectral interferometry

Multi-wavelength digital holographic tomography based on spe...

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

作者： Yu, Lingfeng Chen, Zhongping Univ Calif Irvine Dept Biomed Engn Beckman Laser Inst Irvine CA 92612 USA

ISBN: (纸本)9780819474308

We report a multi-wavelength digital holographic tomography system based on a fiber-based spectral interferometer. Instead of using tunable lasers, a relatively inexpensive broadband source was used as the light source. Hundreds of 2-D "synthesized holograms" (or object wavefields) were obtained by transversal scanning of a probe beam within a few seconds. Holographic images of an object volume were numerically reconstructed from each synthesized hologram and tomographic images were obtained by superposition of all the image volumes.

关键词： Digital holography Tomography Holograms Holography 3D image reconstruction Holographic interferometry Laser beam diagnostics Reconstruction algorithms 3D image processing Interferometry

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Real-time digital design for an optical coherence tomography acquisition and processing system

Real-time digital design for an optical coherence tomography...

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

作者： Ralston, TS Mayen, JA Marks, D Boppart, SA Univ Illinois Beckman Inst Adv Sci & Technol Urbana IL 61801 USA

ISBN: (纸本)0819452327

We present a real-time, multi-dimensional, digital, optical coherence tomography (OCT) acquisition and imaging system. The system consists of conventional OCT optics, a rapid scanning optical delay (RSOD) line to support fast data acquisition rates, and a high-speed A/D converter for sampling the interference waveforms. A 1M-gate Virtex-II field programmable gate array (FPGA) is designed to perform digital down conversion. This is analogous to demodulating and low-pass filtering the continuous time signal. The system creates in-phase and quadrature-phase components using a tunable quadrature mixer. Multistage polyphase finite impulse response (FIR) filtering and down sampling is used to remove unneeded high frequencies. A floating-point digital signal processor (DSP) computes the magnitude and phase shifts. The data is read by a host machine and displayed on screen at real-time rates commensurate with the data acquisition rate. This system offers flexible acquisition and processing parameters for a wide range of multi-dimensional optical microscopy techniques.

关键词： digital signal processing optical coherence tomography Doppler spectroscopic real-time

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3D microscopy with a double-helix point spread function

3D microscopy with a double-helix point spread function

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

作者： Pavani, Sri Rama Prasanna Piestun, Rafael Univ Colorado Dept Elect & Comp Engn Boulder CO 80309 USA

ISBN: (纸本)9780819474308

Double-helix point spread function (DH-PSF) is an engineered three-dimensional (3D) PSF specifically designed for 3D position estimation and imaging. It exhibits two lobes that rotate continuously around the optical axis with propagation. An information theoretical analysis shows that the DH-PSF carries higher and more uniform Fisher Information than the standard PSF through the 3D volume of interest. Experiments with DH-PSF demonstrate nanometer scale position localization accuracies in all three dimensions. Further, a variety of microscopy techniques such as bright-field, dark-field, and fluorescence can be directly transformed into their DH-PSF counterparts by placing an appropriate phase mask in the imaging path. The flexibility and performance make the DH-PSF attractive for 3D position localization in both photon-limited and photon-unlimited regimes.

关键词： three-dimensional microscopy Point spread function engineering Bright-field Dark-field Fluorescence Computational Optical sensing and imaging

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Quasi-confocal fluorescence sectioning with dynamic speckle illumination

Quasi-confocal fluorescence sectioning with dynamic speckle ...

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

作者： Ventalon, Cathie Mertz, Jerome Boston Univ Dept Biomed Engn 44 Cummington St Boston MA 02215 USA

ISBN: (纸本)0819461326

We present a new fluorescence microscopy technique that provides depth discrimination in thick tissue. The technique relies on a simple modification to a conventional wide-field microscope, and consists in illuminating the sample with a sequence of speckle patterns and displaying the RMS or the variance of the resultant sequence of fluorescent images. We demonstrate quasi-confocal optical sectioning with an axial resolution of 5 microns FWHM. The lateral resolution is identical to the widefield microscope, namely 0.6 micron FWHM. images of a mouse brain are compared with standard wide-field images, demonstrating an efficient quasi-sectioning capacity in thick tissue throughout approximatively 100 microns depth. This is achieved because of the high contrast maintained by speckle in a scattering media.

关键词： fluorescence microscopy optical sectioning 3D imaging linear microscopy confocal microscopy speckle structured light illumination thick tissue imaging

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System for fluorescence lifetime imaging microscopy

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Proceedings of SPIE - The International Society for Optical Engineering 1999年 3605卷 55-64页

作者： van Geest, Lambertus K. Boddeke, Frank R. van Dijk, Pieter W. Kamp, Arjen F. van der Oord, Cornelis J.R. Stoop, Karel W.J. Lambert Instruments Netherlands

ISBN: (纸本)0819430757;9780819430755

An affordable, robust and easy-to-use fluorescence lifetime imaging microscopy (FLIM) workstation that is completely automated and does not need any difficult calibration procedure is introduced. The workstation consists of a standard fluorescence microscope, a modulated excitation light source, a camera, a modulation signal generator and acquisition/processing software. An evaluation of the system and its critical components is presented.

关键词： Optical microscopy

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