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

作者： Riza, NA Bokhari, A Nuonics Inc Orlando FL 32817 USA

ISBN: (纸本)0819452327

Ideally, a no-moving parts fast and agile scanning confocal microscope system is required that can produce true realtime 3-D scans with precision and repeatability. In this paper, such agile optical confocal microsopy designs are proposed that enable high speed precise non-invasive 3-D imaging. These compact confocal microscopes can provide real-time pin-point focussed imaging to enable confocal slices in-vivo, thus greatly reducing motion artifacts. These microscopes can be modified into interferometric microscopes for phase contrast imaging. The proposed microscopes can also greatly improve confocal fluorescence imaging as needed for cancer detection. An ultracompact confocal probe tip connected to a single ultra-thin fiber is another design option allowing flexibility for usage in tight cavities.

关键词： confocal microscopy fiber-optics instrument 3-D imaging

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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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Fast IR imaging with sub-wavelength resolution using a transient near-field probe (tipless near-field microscopy)

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Proceedings of SPIE - The International Society for Optical Engineering 1999年 3605卷 31-39页

作者： Palanker, Daniel V. Smith, Todd I. Schwettman, H.Alan Stanford Univ Stanford United States

ISBN: (纸本)0819430757;9780819430755

A technique that enables infrared (IR) imaging with the resolution of a visible microscope is introduced. The technique uses conventional IR and no near-field tips. The resolution of the IR microscope is determined by the dimensions of the transient mirror. To prevent resolution degradation due to diffusion of the photo-excited carriers in the substrate, the probe (IR) pulse duration should not exceed a few tens of picoseconds.

关键词： Optical microscopy

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processing and improvements in dynamic quantitative phase microscope

Processing and improvements in dynamic quantitative phase mi...

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

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

ISBN: (纸本)9780819493583

This paper describes recent research and development related to data processing and imaging performance for a dynamic quantitative phase imaging microscope. This microscope provides instantaneous measurements of dynamic motions within and among live cells without labels or contrast agents. It utilizes a pixelated wire grid polarizer mask in front of the camera sensor that enables simultaneous measurement of multiple interference patterns. Optical path difference (OPD) and optical thickness (OT) data are obtained from phase images. Simulated DIC (gradient) and simulated dark field (gradient magnitude) images can be directly obtained from the phase enabling simultaneous capture of brightfield, phase contrast, quantitative phase, DIC and dark field. The OT is further processed to remove background shapes, and enhance topography. This paper presents a number of different processing routines to remove background surface shape enabling quantification of changes in cell position and volume over time. Data from a number of different moving biological organisms and cell cultures are presented.

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

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Tradeoff between insensitivity to depth-induced spherical aberration and resolution of 3D fluorescence imaging due to the use of wavefront encoding with a radially symmetric phase mask 25

Tradeoff between insensitivity to depth-induced spherical ab...

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

作者： Doblas, Ana Dutta, Ananya Saavedra, Genaro Preza, Chrysanthe Univ Memphis Dept Elect & Comp Engn Memphis TN 38152 USA Univ Valencia Dept Opt E-46100 Burjassot Spain

ISBN: (数字)9781510614840

ISBN: (纸本)9781510614840;9781510614833

Previously, a wavefront encoded (WFE) imaging system implemented using a squared cubic (SQUBIC) phase mask has been verified to reduce the sensitivity of the imaging system to spherical aberration (SA). The strength of the SQUBIC phase mask and, as consequence, the performance of the WFE system are controlled by a design parameter, A. Although the higher the A-value, the more tolerant the WFE system is to SA, this is accomplished at the expense of the effective imaging resolution. In this contribution, we investigate this tradeoff in order to find an optimal A-value to balance the effect of SA and loss of resolution.

关键词： three-dimensional microscopy wavefront encoding point-spread function engineering resolution limit

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Variable-focus microlenses as a potential technology for endoscopy

Variable-focus microlenses as a potential technology for end...

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

作者： Smith, PJ McCabe, EM Taylor, CM Selviah, DR Day, SE Commander, LG Univ Dublin Trinity Coll Dept Phys Dublin 2 Ireland

ISBN: (纸本)081943535X

Using a new device which contains an array of microlenses whose focal lengths can be electrically varied, we have been able to control the input from one microlens to a single mode fibre using an applied voltage. For such a microlens array many closely-spaced focal spots can be generated in parallel, and electrically switched to address, potentially, an array of receiver fibres. We show how the particular switching characteristics of the device, whereby the lenses switch from diverging to converging, serves in turn to disperse light and to focus it into the fibre.

关键词： microlenses endoscopy direct view microscopy

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Photobleaching Property of Confocal Laser Scanning microscopy with Masked Illumination

Photobleaching Property of Confocal Laser Scanning Microscop...

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

作者： Kim, DongUk Moon, Sucbei Song, Hoseong Yang, Wenzhong Kim, Dug Y. Gwangju Inst Sci & Technol Dept Informat & Commun Kwangju 500712 South Korea

ISBN: (纸本)9780819479662

Confocal laser scanning microscopy (CLSM) has become the tool of choice for high-contrast fluorescence imaging in the study of the three-dimensional and dynamic properties of biological system. However, the high cost and complexity of commercial CLSMs urges many researchers to individually develop low cost and flexible confocal microscopy systems. The high speed scanner is an influential factor in terms of cost and system complexity. Resonant galvo scanners at several kHz have been commonly used in custom-built CLSMs. However, during the repeated illumination for live cell imaging or 3D image formation, photobleaching and image distortion occurred at the edges of the scan field may be more serious than the center due to an inherent property (e. g. sinusoidal angular velocity) of the scan mirror. Usually, no data is acquired at the edges due to large image distortion but the excitation beam is still illuminated. Here, we present the photobleaching property of CLSM with masked illumination, a simple and low cost method, to exclude the unintended excitation illumination at the edges. The mask with a square hole in its center is disposed at the image plane between the scan lens and the tube lens in order to decrease photobleaching and image distortion at the edges. The excluded illumination section is used as the black level of the detected signals for a signal quantizing step. Finally, we demonstrated the reduced photobleaching at the edges on a single layer of fluorescent beads and real-time image acquisition without a standard composite video signal by using a frame grabber.

关键词： Photobleaching confocal laser scanning microscopy video-rate 3D image masked illumination

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Effects of specimen morphology on adaptive confocal and multiphoton microscopy

Effects of specimen morphology on adaptive confocal and mult...

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

作者： Booth, Martin J. Wilson, Tony Univ Oxford Dept Engn Sci Parks Rd Oxford OX1 3PJ England

ISBN: (纸本)9780819465566

microscopy of thick biological specimens is often detrimentally affected by specimen-induced aberrations. In the simplest case, these aberrations arise from a refractive index mismatch between the immersion and mounting media. In other situations, the aberrations arise from variations in refractive index within the specimen. These aberrations cause loss of signal and reduced resolution. Aberrations can be corrected using adaptive optics, where a deformable mirror introduces equal but opposite aberrations into the optical path. Aberration correction can be performed by reconfiguring the deformable mirror and using the fluorescence signal as feedback, effectively maximising the fluorescence intensity. However, the degree to which aberrations affect the intensity is related to the distribution of fluorescence in the specimen. Signals from point-like objects are affected more than equivalent signals from from planar or volume objects. We investigate this effect and discuss the implications for adaptive optical microscopy of biological specimens.

关键词： confocal microscopy multiphoton microscopy aberrations adaptive optics

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Experimental analysis of focal fields in laser scanning fluorescence stereomicroscopy

Experimental analysis of focal fields in laser scanning fluo...

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

作者： Yang, Yanlong Peng, Tong Lei, Ming Li, Runze Zhou, Xing Wu, Di Yao, Baoli Ye, Tong Chinese Acad Sci Xian Inst Opt & Precis Mech State Key Lab Transient Opt & Photon Xian Shaanxi Peoples R China Clemson Univ Ctr Optic Mat Sci & Engn Technol Dept Bioengineering Clemson SC 29631 USA

ISBN: (纸本)9780819498625

Bessel beams have been proved to have the ability to extend the depth of focus in fluorescence microscopy. But the depth discrimination was not investigated thoroughly. Following our previous work(2), we investigated focal fields of Bessel-Gauss beams at different scanning angles. We found that the central focusing lines were tilted differently at different scanning angles. This effect manifests the ability of the true perspective view in the fluorescence stereomicroscopy.

关键词： Bessel-Gauss beams extended depth of field stereomicroscopy laser scanning three-dimensional imaging volume imaging

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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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