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

作者： Facomprez, Aurelie Beaurepaire, Emmanuel Debarre, Delphine Ecole Polytech ParisTech Lab Opt & Biosci CNRS F-91128 Palaiseau France INSERM F-91128 Palaiseau France

ISBN: (纸本)9780819488701

We investigate the parameters governing the accuracy of correction in modal sensorless adaptive optics for microscopy. In this paper we focus on the case of two-photon excited fluorescence. Using analytical, numerical and experimental results, we show that using a suitable number of measurements, accurate correction can be achieved for up to 2 rad rms initial aberrations even without optimisation of the correction modes. We demonstrate that this correction can be achieved using low light levels to minimise photobleaching and toxicity, and we provide examples of such optimised correction.

关键词： Adaptive optics aberration correction multiphoton microscopy nonlinear microscopy

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An optical technique for fast focusing applied to high-aperture microscopy

An optical technique for fast focusing applied to high-apert...

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

作者： Botcherby, Edward Juskaitis, Rimas Booth, Martin Wilson, Tony Univ Oxford Dept Engn Sci Parks Rd Oxford OX1 3PJ England

In this paper we describe a simple method of optical refocusing for high numerical aperture imaging systems. As the first stage of this new method satisfies both the sine and Herschel conditions of geometric optics, i... 详细信息

ISBN: (纸本)9780819465566

关键词： optical refocusing sine condition Herschel condition confocal imaging

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A comparison of methods for optical sectioning using structured illumination microscopy

A comparison of methods for optical sectioning using structu...

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

作者： Thomas, Benjamin Momany, Michelle Kner, Peter Univ Georgia Coll Engn Athens GA 30602 USA Univ Georgia Dept Plant Biol Athens GA 30602 USA

ISBN: (纸本)9780819493583

Structured Illumination microscopy is a simple and effective method to remove out-of-focus light in widefield fluorescence microscopy. Neil et al. originally proposed a simple square-law method for calculating the optically sectioned image from the three raw images with the structured illumination pattern super-imposed. However, the Neil method does not make the most efficient use of the three raw images. The three structured illumination images can also be used to separate three copies of the image covering shifted regions of frequency space in a similar manner to that developed by Gustafsson et al. These can then be combined using a generalized Wiener filter to create an image with a well-behaved optical transfer function in which the missing cone has been filled in, providing optical sectioning. Here, we compare the Neil and Gustafsson methods and show that the Gustafsson method provides an image with higher fidelity and a better Signal to Noise Ratio (SNR) at low photon counts. We apply the two methods to images of fluorescent beads and GFP labeled septins in Aspergillus nidulans.

关键词： fluorescence microscopy structured illumination microscopy three-dimensional imaging bioimaging

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Nano-scale optical imaging technique

Nano-scale optical imaging technique

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

作者： Wu, S Tang, YC PEER Ctr Covina CA 91722 USA

ISBN: (纸本)0819452327

The optical resolution limits the observation of fine features with size below diffraction limit, e.g. 500nm. We propose and demonstrate a technique that uses collimated laser scattering to characterize such features. We use a new illumination technique that let us image bio-molecules down to 50nm with optical microscopy. We observed vesicles with size down to 50nm and visualize the flow of such tiny molecules in the live cells. We could also characterize the detailed size and molecular weight of such particles.

关键词： single particle laser scattering microscopy

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microscopy using an interference pattern as illumination source

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

作者： Juskaitis, R Neil, MAA Wilson, T Univ Oxford Dept Engn Sci Oxford OX1 3PJ England

ISBN: (纸本)0819427004

We describe a method of obtaining optically sectioned fluorescence images in a widefield conventional microscope by interfering two beams on an object so as to illuminate it with a single spatial frequency fringe pattern. images taken at three spatial positions of the fringe pattern are processed in real time to produce optically sectioned images which are substantially similar to those obtained with confocal microscopes.

关键词： 3D microscopy confocal microscopy fluorescence microscopy

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A compact polarization converter to observe molecular orientation

A compact polarization converter to observe molecular orient...

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

作者： Hashimoto, Mamoru Yoshiki, Keisuke Kanamaru, Ryosuke Hashimoto, Nobuyuki Araki, Tsutomu Osaka Univ Grad Sch Engn Sci 1-3 Machikaneyama Toyonaka Osaka 5608531 Japan Citizen Watch Co Ltd MHT R&D Dept Tokorozawa Saitama 3598511 Japan

ISBN: (纸本)9780819465566

We developed a compact polarization-converter using two liquid-crystal spatial-light-modulators with eight electrodes. The converter converted a linearly polarized beam to two orthogonal linearly polarized beams and a radially polarized beam, and the direction of the electric filed at the focal point were controlled three-dimensionally. We constructed a second-harmonic-generation microscope using the polarization-converter to observe three-dimensional molecular orientation and demonstrated the detectability of molecular orientation.

关键词： molecular orientation nonlinear microscopy second-harmonics-generation pupil filtering radially polarized beam

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A new expanded point information content design approach for 3D live-cell microscopy at video rates

A new expanded point information content design approach for...

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

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)9780819493583

Through a combination of optical design and algorithm development, a new expanded point information content ( EPIC) microscope has been developed that is capable of extending the depth of field while simultaneously super locating the depth position of complex biological objects to within an accuracy of 75 nm. The data is then combined to form 3D animations of live-cell biological specimens. This is accomplished without the need to acquire multi-focal image stacks and is thus well suited for high-speed imaging.

关键词： 4D microscopy PSF engineering extended depth of field quantitative depth ranging computational optics live-cell imaging fluorescence microscopy

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three-dimensional reconstruction and analysis of gastric malignancies by electronic slides of consecutive sections and virtual microscopy

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Proceedings of SPIE - The International Society for Optical Engineering 1999年 3605卷 190-199页

作者： Molnar, Bela Papik, Kornel Tagscherer, Attila Csendes, Gabor Varga, Sebestyen V. Tulassay, Zsolt Schaefer, Rainer Mahoney, Walt Semmelweis Univ Budapest Hungary

ISBN: (纸本)0819430757;9780819430755

A study was carried out focusing on the application of an electronic slide and a software simulated virtual microscope to histological analysis. The electronic slide was developed using a fully computer controlled microscope, video camera and an Intel Pentium II based PC. A program called Pyramid was developed for the scanning of an entire slide. Autofocusing, image reduction and frame joining algorithms were implemented in the virtual microscope application. Results confirm that the electronic slide and the software contribute to a more efficient, convenient histological analysis.

关键词： Optical microscopy

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Fast 3D blind deconvolution of even point spread functions

Fast 3D blind deconvolution of even point spread functions

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

作者： Yagle, AE Shah, S Univ Michigan Dept EECS Ann Arbor MI 48109 USA

ISBN: (纸本)0819452327

3-D mages obtained from optical sectioning microscopy are usually degraded by a point-spread function that is known to be an even function but is otherwise only approximately known, or even entirely unknown. We present a new algorithm for 3-D blind deconvolution of even point-spread functions that is both fast and (in the absence of noise) exact. Fourier transforms decouple the problem into 2-D, then 1-D blind deconvolution problems, greatly increasing computational speed. Numerical simulations demonstrate that the blind algorithm seems to perform both faster and more accurately than the non-blind iterative Lucy-Richardson algorithm.

关键词： Deconvolution Point spread functions 3D image processing Signal to noise ratio Convolution microscopy Fourier transforms Confocal microscopy image filtering Matrices

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Confocal scanning microscopy with multiple optical probes for high speed measurements and better imaging - art. no. 686115

Confocal scanning microscopy with multiple optical probes fo...

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

作者： Chun, Wanhee Lee, SeungWoo Gweon, Dae-Gab Korea Adv Inst Sci & Technol Div Mech Engn Nano Opto Mechatron Lab Taejon 305701 South Korea

ISBN: (纸本)9780819470362

Confocal scanning microscopy (CSM) needs a scanning mechanism because only one point information of specimen can be obtained. Therefore the speed of the confocal scanning microscopy is limited by the speed of the scanning tool. To overcome this limitation from scanning tool we propose another scanning mechanism. We make three optical probes in the specimen under confocal condition of each point. three optical probes are moved by beam scanning mechanism with shared resonant scanning mirror (RM) and galvanometer driven mirror (GM). As each optical probe scan allocated region of the specimen, information from three points is obtained simultaneously and image acquisition time is reduced. Therefore confocal scanning microscopy with multiple optical probes is expected to have three times faster speed of the image acquisition than conventional one. And as another use, multiple optical probes to which different light wavelength is applied can scan whole same region respectively. It helps to obtain better contrast image in case of specimens having different optical characteristics for specific light wavelength. In conclusion confocal scanning microscopy with multiple optical probes is useful technique for views of image acquisition speed and image quality.

关键词： confocal scanning microscopy multi point scanning multiple optical probes high speed measurement better image quality color imaging

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