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

作者： Miao, Qin Yu, Julia Meyer, Michael G. Rahn, J. Richard Neumann, Thomas Nelson, Alan C. Seibel, Eric J. Univ Washington Dept Bioengn Seattle WA 98195 USA VisGate Inc Gig Harbor WA 98335 USA Univ Washington Dept Engn Mech Seattle WA 98195 USA

ISBN: (纸本)9780819479662

A dual-modal optical projection tomography microscope (OPTM) is presented, which produces three-dimensional images of single cells with isometric high resolution both in fluorescence and absorption mode. Depth of field of a high numerical aperture objective is extended by scanning the focal plane through the sample in order to enable reconstruction by back-projection method. Cells are fixed, stained, and mixed with optical gel and injected into the capillary for imaging. Combining absorption and fluorescence mode allows us to image different aspects of the disease process. images of cells stained with both hematoxylin and fluorescence probes are shown. Registrations between two modes are discussed.

关键词： three-dimensional microscopy Tomographic imaging Fluorescence biomarker imaging Coregistration

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High speed phase distortion measurement and compensation for focusing in space and time

High speed phase distortion measurement and compensation for...

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

作者： Fiolka, Reto Cui, Meng Howard Hughes Med Inst Ashburn VA 20147 USA

ISBN: (纸本)9780819493583

Random scattering and aberrations severely limit the imaging depth in optical microscopy. We introduce a rapid, parallel wavefront compensation technique that efficiently compensates even highly complex phase distortions. Using coherence gated backscattered light as a feedback signal, we focus light deep inside highly scattering brain tissue. We demonstrate that the same wavefront optimization technique can also be used to compensate spectral phase distortions in ultrashort laser pulses using nonlinear iterative feedback. We can restore transform limited pulse durations at any selected target location and compensate for dispersion that has occurred in the optical train and within the sample.

关键词： Fluorescence microscopy adaptive optics imaging through turbid media ultrafast optics

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Simultaneous quantitative depth mapping and extended depth of field for 4D microscopy through PSF engineering

Simultaneous quantitative depth mapping and extended depth o...

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

作者： Zahreddine, Ramzi N. Cormack, Robert H. Cogswell, Carol J. Univ Colorado Dept Elect Comp & Energy Engn Boulder CO 80309 USA

ISBN: (纸本)9780819488701

An extended depth of field (EDF) microscope that allows for quantitative axial positioning has been constructed. Past work has shown that EDF microscopy allows for features in varying planes to appear sharply focused simultaneously, however an inherent consequence of this is that depth information is lost. Here, a specifically engineered phase plate is used to create a point spread function (PSF) that contains both of the necessary attributes for extended depth of field and quantitative depth mapping. A two-camera solution is used to separate and capture the information for individualized post processing. The result is a microscope that can serve as an essential tool for full 3D, real-time imaging.

关键词： 4D microscopy PSF engineering extended depth of field quantitative depth ranging

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High-contrast 3D microscopic imaging of deep layers in a biological medium

High-contrast 3D microscopic imaging of deep layers in a bio...

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

作者： Faridian, Ahmad Pedrini, Giancarlo Osten, Wolfgang Univ Stuttgart Inst Tech Opt SCoPE Pfaffenwaldring 9 D-70569 Stuttgart Germany

ISBN: (纸本)9780819498625

Multilayer imaging of biological specimens is a demanding field of research, but scattering is one of the major obstacles in imaging the internal layers of a specimen. Although in many studies the biological object is assumed to be a weak scatterer, this condition is hardly satisfied for sub-millimeter sized organisms. The scattering medium is inhomogeneously distributed inside the specimen. Therefore, the scattering which occurs in the upper layers of a given internal layer of interest is different from the lower layers. That results in a different amount of collectable information for a specific point in the layer from each view. An opposed view dark-field digital holographic microscope (DHM) has been implemented in this work to collect the information concurrently from both views and increase the image quality. Implementing a DHM system gives the possibility to perform digital refocusing process and obtain multilayer images from each side without depth scanning of the object. The results have been presented and discussed here for a Drosophila embryo.

关键词： Digital holographic microscopy Multilayer imaging three-dimensional microscopy image enhancement image fusion

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Coherence measurements applied to critical and Kohler vortex illumination

Coherence measurements applied to critical and Kohler vortex...

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

作者： Brown, Dean P. Brown, Thomas G. Univ Rochester Inst Opt Rochester NY 14627 USA

ISBN: (纸本)9780819474308

Both coherence and polarization play an important role in microscopy. It was long ago established that, in the context of scalar treatments, Kohler and critical illumination produce equivalent coherence functions in an object plane. This paper examines this equivalence in the context of polarization vortex illumination. Using a reversed-wavefront Young interferometer (RWYI), we show measurements of the coherence and correlation properties of the output illumination plane of various illumination systems. We compare the coherence properties of critical and Kohler vortex illumination schemes and look at the effects these properties have in an imaging system.

关键词： Coherence Polarization Vortex Partial Coherent Imaging Critical Illumination Kohler Illumination Polarization microscopy

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Using out-of-focus light to improve image acquisition time in confocal microscopy

Using out-of-focus light to improve image acquisition time i...

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

作者： Davis, BJ Karl, WC Goldberg, BB Swan, AK Ünlü, MS Boston Univ Dept Elect & Comp Engn Boston MA 02215 USA

ISBN: (纸本)0819456756

Light that would typically be discarded at a confocal microscope's detector pinhole will be collected and processed to allow a reduced spatial sampling rate and thus an improved image acquisition time. It is shown that collecting and appropriately processing the out-of-focus light will allow an axial sampling rate below that specified by the Nyquist criterion. To achieve this, a central detector pinhole and a number of out-of-focus regions are collected concurrently. This corresponds to imaging through several different channels, with differing point spread functions, in parallel. Since the spatial sampling rate is below the Nyquist frequency, aliasing occurs in the data from each of the channels. However, since the point spread functions are different, the aliasing effects are different in each channel. This allows the ensemble of aliased images to be processed into a sin-le dealiased and deconvolved image. This potential utility of out-of-focus light is demonstrated through simulated examples for differing collection schemes and scanning rates. Results are shown for under-sampling by up to a factor of four. Collecting the out-of-focus light also improves instrument collection efficiency.

关键词： confocal microscopy image acquisition time sampling rate aliasing out-of-focus light confocal pinhole high speed microscopy confocal collection efficiency

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A study for accelerating the speed of all-in-focus image processing 27

A study for accelerating the speed of all-in-focus image pro...

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

作者： Wang, Lihui Tabata, Satoshi Takeuchi, Hirotoshi Ishikawa, Masatoshi Guangdong Acad Sci Guangdong Inst Semicond Ind Technol 363 Changxing Rd Guangzhou 510650 Peoples R China Univ Tokyo Grad Sch Informat Sci & Technol Bunkyo Ku 7-3-1 Hongo Tokyo 1138656 Japan

ISBN: (纸本)9781510632547

To capture an all-in-focus and 3D depth image, shape from focus method is widely used. The phase accuracy of the image candidates should be processed and adjusted beforehand Phase only correction method was employed in this work and to accelerate the processing speed, the image processing by Fast Fourier Transform (FFT) was optimized Meanwhile, the processing task was assigned in several parallel threads so that the performance would be improved. The method was used on a variable focus imaging system, and, as a result, the processing speed was improved to around 2.5-fps.

关键词： all-in-focus shape from focus phase-only correction method 3D imaging variable focus system

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Polarised light microscopy

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

作者： Torok, P Higdon, PD Wilson, T Univ Oxford Dept Engn Sci Oxford OX1 3PJ England

ISBN: (纸本)0819427004

In this work we discuss the role of polarisation in confocal microscopy. The effect of the optical system on light polarisation is analysed in the absence of a specimen. We also present numerical results on confocal microscopes imaging small dielectric scatterers and suggest an optimum pinhole size for crossed linear polars. Finally, imaging of large spherical scatterers and contrast optimisation of the optical system is discussed.

关键词： confocal microscopy polarised light vectorial theory high aperture theory

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Transparent three-dimensional microstructures for trapping of living cells

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Proceedings of SPIE - The International Society for Optical Engineering 1999年 3605卷 241-249页

作者： Otte, K. Zimmer, K. Zeitschel, U. Hirsch, D. Reichenbach, A. Bigl, F. Bigl, V. Inst fuer Oberflaechenmodifizierung e.V. Leipzig Germany

ISBN: (纸本)0819430757;9780819430755

A combination of e-beam lithography (EBL), laser ablation, reactive ion beam etching (RIBE) and different wet etching techniques was used to fabricate different transparent three-dimensional structures in a three-layer system (Si3N4, SiO2, Si3N4) on quartz specially sized for the trapping of living neural cells under physiological conditions. The resulting structures show potential for trapping of cells, the stabilization of the cell membrane and the precise placement of the cells for a multimode of biological investigations.

关键词： Atomic force microscopy

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Improved image-forming optics for transmission optical projection tomography

Improved image-forming optics for transmission optical proje...

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

作者： Wang, Yi Wang, Ruikang Oregon Hlth & Sci Univ Dept Biomed Engn Beaverton OR 97006 USA Foshan Univ Dept Photoelect & Phys Foshan 528000 Peoples R China

ISBN: (纸本)9780819465566

We demonstrate an improved image-forming optics for transmission optical projection tomography (TOPT), with which the parallel integral throughout an object can be obtained. This is performed by introducing a shutter with an appropriate diameter at the back focus of the objective lens. We evaluate and compare the performances of the improved and the conventional TOPT systems with different parameters to obtain the optimal configuration. The optimal reconstruction is achieved by the improved configuration with alpha = 0.8 degrees similar to 1.6 degrees, and the spatial resolution reaches 25 mu n. The optimal configuration is validated by TOPT of a phantom sample and a five days chick embryo.

关键词： optical projection tomography point spread function parallel integral

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