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Confocal photoluminescence microscopy in II-VI materials: annealing and degradation dynamics

Proceedings of SPIE - The International Society for Optical Engineering 1999年 3605卷 65-72页

作者： McCabe, E.M. Jordan, C. Fewer, D.T. Donegan, J.F. Taniguchi, S. Hino, T. Nakano, K. Ishibashi, A. Uusimaa, P. Pessa, M. Trinity Coll Dublin Dublin Ireland

A new approach to imaging degradation and annealing in situ in a range of II-VI materials is presented. The approach uses confocal photoluminescence imaging in a confocal microscope. Using this imaging technique, it i... 详细信息

ISBN: (纸本)0819430757;9780819430755

关键词： Optical microscopy

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Near-field optical and atomic force constraints for super-resolution 3D deconvolution in far-field optical microscopy

Near-field optical and atomic force constraints for super-re...

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

作者： Axelrod, N Radko, A Ben-Yosef, N Khatchatouriants, A Treinin, M Lewis, A Hebrew Univ Jerusalem Dept Appl Phys IL-91904 Jerusalem Israel

ISBN: (纸本)081943535X

We demonstrate that near-field optical and atomic force microscopy data can be used for super-resolution three-dimensional (3-D) image restoration in optical sectioning fluorescence microscopy. A crucial feature in this approach is the full integration of such data sets with digital far-field images. The scanned probe data is used to provide modalities for boundary constraints which define surface optical information and spatial domains of optical alterations in a sample with a spatial precision that has been unachievable in the past. A restoration algorithm that can use such a complex of data for 3-D image deconvolution is presented. It uses a Tikhonov-Miller regularization scheme and allows for the imposition of different types of constraints to obtain super-resolution deconvolved images. Performance was tested by using simulated 3-D imaging. An example is given of the restoration of a 3-D wide field optical image of a biological specimen in the presence of atomic force constraints.

关键词： image restoration deconvolution algorithm constraints probe microscopy NSOM super-resolution

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Extended depth-of-field microscopy

Extended depth-of-field microscopy

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

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

ISBN: (纸本)9780819479662

In this paper, we present details of a scanning two-photon fluorescence microscope we have built with a near-isotropic scan rate. This means that the focal spot can be scanned at high speed along any direction in the specimen, without introducing systematic aberrations. We present experimental point spread function measurements for this system using an Olympus 1.4 NA 60X oil immersion lens that demonstrates an axial range of operation greater than 70 mu m. We give details of a novel actuator device used to displace the focusing element and demonstrate axial scan responses up to 3.5 kHz. Finally, we present an application of this system in liquid crystal research to image the dynamic response of a nematic device during switching. Information about the director field at different levels in the device can be inferred from images acquired with a temporal resolution of 2.5 ms.

关键词： optical refocusing two-photon microscopy

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FPGA Implementation of Spectral Fusing Gabor Domain Optical Coherence microscopy 27

FPGA Implementation of Spectral Fusing Gabor Domain Optical ...

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

作者： Meemon, Panomsak Lenaphet, Yutana Saetiew, Jadsada Suranaree Univ Technol Sch Phys Inst Sci Nakhon Ratchasima 30000 Thailand Suranaree Univ Technol Sch Elect Engn Inst Engn Nakhon Ratchasima 30000 Thailand

ISBN: (纸本)9781510632547

Gabor domain optical coherence microscopy (GD-OCM) is an emerging tomographic imaging technique that overcome the depth of focus (DOF) limit of high numerical objective lens by acquiring multiple cross-section images corresponded with different focal planes of the objective along an axial direction and combining them to form an invariant high resolution cross-section image. We previously reported an alternative processing method of GD-OCM called spectral fusing GD-OCM (SF-GD-OCM). SF-GD-OCM extracts in-focus information directly from acquired spectral interference signals. The advantage of SF-GD-OCM over the original GD-OCM is that the Gabor filtering and fusing can be implemented in an acquisition hardware instead of a host computer. Here, we have investigated the implementation of SF-GD-OCM by using FPGA frame grabber. The experimental setup of the acquisition system as well as the implementation of the spectral Gabor fusion in FPGA device have been developed and verified. Results show that the data processing of SF-GD-OCM in FPGA enabled acquisition device is possible and could lead to improvement in overall imaging speed of GD-OCM.

关键词： Gabor domain optical coherence microscopy Gabor fusion technique FPGA Extended depth of field

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Structured illumination microscopy using photoswitchable fluorescent proteins

Structured illumination microscopy using photoswitchable flu...

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

作者： Hirvonen, Liisa Mandula, Ondrej Wicker, Kai Heintzmann, Rainer Kings Coll London Randall Div Cell & Mol Biophys London SE1 1UL England

ISBN: (纸本)9780819470362

In fluorescence microscopy the lateral resolution is limited to about 200 nm because of diffraction. Resolution improvement by a factor of two can be achieved using structured illumination, where a fine grating is projected onto the sample, and the final image is reconstructed from a set of images taken at different grating positions. Further resolution improvement can be achieved by saturating the transitions involved in fluorescence emission. Recently discovered photoswitchable proteins undergo transitions that are saturable at low illumination intensity. Combining this concept with structured illumination, theoretically unlimited resolution can be achieved, where the smallest resolvable distance will be determined by signal-to-noise ratio. This work focuses on the use of the photoswitchable protein Dronpa with structured illumination to achieve nanometre scale resolution in fixed cells.

关键词： structured illumination nonlinear excitation photoswitching Dronpa

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Fast scanning confocal microscopy with a rotary mirror array

Fast scanning confocal microscopy with a rotary mirror array

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

作者： Chen, NG Zhu, Q Univ Connecticut Dept Elect & Comp Engn Storrs CT 06269 USA

ISBN: (纸本)0819447641

We have developed a novel confocal microscopy system, which can perform both axial and lateral scanning at an ultra high speed. A rotary mirror array (RMA) consists of 36 small mirrors deployed on the surface of a round plate with a small tilting angle and a discrete rotational symmetry. When the RMA rotates at a constant speed, the focal point is scanned inside a sample periodically and linearly. We have demonstrated a scanning rate of 2400 lines per second, which can be readily improved to more than 20000 lines per second. Our scheme can also be used in a fast scanning multi-photon microscope.

关键词： rotary mirror array fast scanning confocal microscopy optical coherence microscopy

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A wide-field microscopy technique using a linear image sensor for obtaining 3D images

A wide-field microscopy technique using a linear image senso...

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

作者： Macedo, MP Barata, AJ Fernandes, AG Correia, CM Inst Engn Coimbra Dept Phys & Math P-3030 Coimbra Portugal

ISBN: (纸本)0819456756

three-dimensional (3D) microscopy is a huge field that includes several microscopy techniques. Among these techniques the confocal microscopy is widely known and used due to its improved axial resolution or optical sectioning ability. However its high cost and image acquisition time as well as low signal-to-noise ratio are important drawbacks. Other techniques use wide-field structured illumination to get depth information from the sample. The detectors in commercial microscopes are normally of the point detector type (PMTs) in confocal and area sensors (CCDs) in widefield microscopy. Examples of microscopes that take advantage of lower cost and faster readout cycles of linear image sensors are very uncommon. The purpose of this work is to develop a low-cost microscopy technique for obtaining 3D images with lower acquisition time and higher signal-to-noise ratio (SNR) than in classical confocal microcopy. It uses a linear CMOS image sensor and specific reconstruction algorithms will be able to extract 3D information from the distribution of light intensity collected. The sensor readout circuitry has been developed and tests are currently running in a laboratory prototype in reflection mode with an epi-illuminated configuration of scanning-stage type. The light source is a commercial incandescent lamp with regulated intensity. For the development, test and optimization of the reconstruction algorithms the object is mounted on a 3-axis translation stage to scan the object in three perpendicular directions. Contrast and resolution results obtained using a resolution test slide and preliminary images of integrated circuit (IC) bonding are presented.

关键词： 3D microscopy confocal microscopy reconstruction algorithms optical sectioning linear sensor

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Phase retrieval of widefield microscopy point spread functions

Phase retrieval of widefield microscopy point spread functio...

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

作者： Hanser, BM Gustafsson, MGL Agard, DA Sedat, JW Univ Calif San Francisco Grad Grp Biophys San Francisco CA 94143 USA

ISBN: (纸本)0819439398

The three-dimensional imaging properties of a light microscope are traditionally described through an intensity point spread function (PSF) or its Fourier transform, the optical transfer function (OTF). However, the imaging properties can be more compactly described by a two-dimensional generalized pupil function. Use of the pupil function allows easy modification of an observed PSF to introduce known aberrations, a task that is much more difficult when using a PSF or OTF. Unfortunately, it is not straightforward to determine the complex-valued pupil function from the measured intensity PSF, because of the lack of phase information. This is the problem of phase retrieval. Several phase retrieval algorithms have been developed for two-dimensional imaging in astronomy. We have modified one such algorithm to be appropriate for the high-aperture, non-paraxial case of high resolution light microscopy. PSFs reconstructed from phase-retrieved pupil functions, modified by calculated aberrations closely reproduce the features of measured, aberrated PSFs.

关键词： phase retrieval Point Spread Function (PSF) wide-field microscopy fluorescence microscopy Optical Transfer Function (OTF) phase diversity optical microscopy pupil function

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QSIP: Phase imaging made possible in a bright field microscope

QSIP: Phase imaging made possible in a bright field microsco...

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

作者： Pavani, Sri Rama Prasanna Libertun, Ariel R. King, Sharon V. Cogswell, Carol J. Univ Colorado Dept Elect & Comp Engn Micro Opt Imaging Syst Lab Boulder CO 80309 USA

ISBN: (纸本)9780819470362

Quantitative structured-illumination phase microscopy (QSIP) uses a conventional bright field microscope to quantitatively measure the optical path length profiles of homogenous phase-only objects. The illumination in QSIP is structured with a predetermined pattern by placing an amplitude mask in the field diaphragm of the microscope. From the image of the amplitude mask, a numerical algorithm implementing a closed form analytical solution calculates the object's optical path length profile. In this paper, we investigate the accuracy of the numerical algorithm used and show that it can be made arbitrarily accurate by using numerical optimization. We then analyze the effect of the system's numerical aperture (NA), and show that QSIP can be used with a wide range of NAs for objects with small phase gradients, and can be used with relatively lower NAs for objects with large phase gradients.

关键词： phase computational imaging structured illumination bright field microscopy metrology

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A phase-shifting DIC technique for measuring 3D phase objects: Experimental verification

A phase-shifting DIC technique for measuring 3D phase object...

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

作者： King, SV Cogswell, CJ Univ Colorado Dept Elect & Comp Engn Boulder CO 80309 USA

ISBN: (纸本)0819452327

Experimental verification of our previously proposed linear phase imaging technique for differential interference contrast microscopy (DIC) microscopy is presented. This technique first applies phase-shifting methods to DIC to acquire linear phase gradient images in two orthogonal directions. A special Fourier integration algorithm is then applied to the combined phase gradient images to create a single linear phase image in which intensity is proportional to phase. This overcomes the limitations of traditional DIC, which cannot accurately measure the phase (i.e. refractive index or thickness) of embedded 3D phase objects. The linear phase imaging technique is implemented using a standard DIC microscope altered to allow controlled phase shifting, a low noise CCD camera, and post-processing in Matlab. The results presented confirm the linear proportionality of intensity to phase in these images.

关键词： DIC phase phase gradient phase imaging spiral phase interference microscopy

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