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检索条件"任意字段=Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing IX"

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Evaluation of tile artifact correction methods for multiphoton microscopy mosaics of whole-slide tissue sections 29

Evaluation of tile artifact correction methods for multiphot...

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

作者： Knapp, Thomas Lima, Natzem Duan, Suzann Merchant, Juanita L. Sawyer, Travis W. Univ Arizona Dept Biomed Engn Tucson AZ 85721 USA Univ Arizona Wyant Coll Opt Sci Tucson AZ 85721 USA Univ Arizona Coll Med Tucson AZ 85724 USA

ISBN: (纸本)9781510648043;9781510648036

Multi-photon microscopy (MPM) is a useful biomedical imaging tool due, in part, to its capabilities of probing tissue biomarkers at high resolution and with depth-resolved capabilities. Automated MPM tile scanning allows for whole-slide image acquisition but suffers from tile-stitching artifacts that prevent accurate quantitative data analysis. We have investigated a variety of post-processing artifact correction methods using imageJ macros and custom Python/MATLAB code and present a quantitative and qualitative comparison of these methods using whole-slide MPM autofluorescence images of human duodenal tissue. image quality is assessed via evaluation of artifact removal compared to the calculated mean square error (MSE), peak signal-to-noise ratio (PSNR), and structural similarity index (SSIM) of the processed image and its raw counterpart. Consideration of both quantitative and qualitative results suggest a combination of flat-field based correction and frequency filtering processing steps provide improved artifact correction when compared to each method used independently to correct for tiling artifacts of tile-scan MPM images.

关键词： Multiphoton microscopy image processing artifacts whole slide multiwavelength tiling

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Quantifying melanin distribution using pump-probe microscopy and a 2D morphological autocorrelation transformation for melanoma diagnosis

Quantifying melanin distribution using pump-probe microscopy...

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

作者： Robles, Francisco E. Wilson, Jesse W. Warren, Warren S. Duke Univ Dept Chem Durham NC 27708 USA

ISBN: (纸本)9780819498625

Pump-probe microscopy is a quantitative molecular imaging technique that yields diagnostically relevant information from endogenous pigments, like melanin, by probing their ultrafast photodynamic properties. Previously, the method was applied to image thin, pigmented, cutaneous samples at different stages of melanoma, and results have shown a correlation between melanin photodynamic behavior and malignancy. Here, we add to the diagnostic power of the method by applying principles of mathematical morphology to parameterize melanins' image structure. Along with bulk melanin chemical information, results show that this method can differentiate invasive melanomas from non-invasive and benign lesions with high sensitivity and specificity (92.3% and 97.5%, respectively, with N = 53). The mathematical method and the statistical analysis are described in detail and results from cutaneous and ocular conjuctival melanocytic lesions are presented.

关键词： Melanoma microscopy Diagnostics Molecules Chemical analysis Modulation Optical parametric oscillators Statistical analysis Molecular imaging Ultrafast imaging

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Fluorescence microscopy with a coded aperture snapshot spectral imager

Fluorescence microscopy with a coded aperture snapshot spect...

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

作者： Fernandez, Christy A. Wagadarikar, Ashwin Brady, David J. McCain, Scott C. Oliver, Tim Duke Univ Fitzpatrick Inst Photon Durham NC 27708 USA Applied Quantum Technol Durham NC 27707 USA Duke Univ Dept Cell Biol Durham NC 27710 USA

ISBN: (纸本)9780819474308

This paper describes the application of a dual-dispersive (DD) coded aperture snapshot spectral imager (CASSI) to fluorescence microscopy. CASSI records an interleaved spatially varying, spectrally filtered map of an object onto a two-dimensional (2D) focal plane. Using a compressive sensing framework, the spectrally encoded 2D scene is reconstructed into a three-dimensional (3D) data cube. CASSI also records a 3D dataset at video rate making it suitable for dynamic cellular imaging. A convex optimization technique combining least squares QR factorization with a total variance constraint is used to reconstruct the image. The system records 32 spectral channels that span the spectral range between 450nm and 750nm with 10nm spectral resolution for any pixel in a 2D image. This paper illustrates the application of CASSI imaging to fluorescence microscopy applications. We report on the reconstruction of fluorescent microspheres used in fluorescence microscopy as calibration standards. images are compared with a multi-spectral confocal system.

关键词： spectral imaging microscopy fluorescence optical systems

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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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Time-resolved confocal microscopy of cryogenic processes in biological tissues

Time-resolved confocal microscopy of cryogenic processes in ...

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

作者： Schellenberg, M. Bergmann, T. Peev, E. Kloster, M. Napier, J. Neu, W. Univ Appl Sci Emden Leer Dept Nat Sci Constantiapl 4 D-26723 Emden Germany Univ Appl Sci Giessen Friedberg Inst Medizin Phys & Strahlenschutz D-35390 Giessen Germany

ISBN: (纸本)9780819484413

Cryogenic procedures are fundamental tools in modern biology, e. g. for conservation or purification of biological materials. The processes occurring in biological cells and tissues during freezing and thawing are subject to ongoing research. Optimization of cell survival rates demands the development and evaluation of exactly defined temperature profiles. 4D-DMD-microscopy is capable of imaging these highly dynamic processes with high spatial and temporal resolution, utilizing well established staining procedures for differentiating structures of interest.

关键词： two-photon DMD confocal microscopy cryogenic process 4D-microscopy cryomicroscopy

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Adapting phasor analysis for nonlinear pump-probe microscopy

Adapting phasor analysis for nonlinear pump-probe microscopy

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

作者： Robles, Francisco E. Wilson, Jesse W. Fischer, Martin C. Warren, Warren S. Duke Univ Dept Chem Durham NC 27708 USA

ISBN: (纸本)9780819493583

Phasor analysis has become a powerful tool for examining signals in fluorescence life-time microscopy (FLIM), where the analysis provides a fast, robust and intuitive means of separating different fluorescent species and mixtures thereof. In this work we adapt this analysis for pump-probe microscopy, a method that provides molecular contrast of pigmented samples by probing their excited state dynamic properties. The bipolar nature of the pump-probe signals presents important differences in the resulting phasors compared to FLIM-here, we discuss these differences and describe the behavior of bipolar signals in phasor analysis. Results show that this method is indeed able to separate multiple molecular species of interests and allows facile assessment of pigment chemistry and its distribution in samples.

关键词： microscopy Molecules Terahertz radiation Signal analyzers Statistical analysis Chemistry Fluorescence lifetime imaging Biological research Laser beam diagnostics Modulation

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Comparison of computational methods developed to address depth-variant imaging in fluorescence microscopy

Comparison of computational methods developed to address dep...

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

作者： Rahman, Muhammad Mizanur Schaefer, Lutz H. Schuster, Dietwald Preza, Chrysanthe Univ Memphis Dept Elect & Comp Engn Memphis TN 38152 USA Adv Imaging Mathodol Consultat Ontario ON Canada Univ Appl Sci Dept Math Regensburg Germany

ISBN: (纸本)9780819493583

In three-dimensional microscopy, the image formation process is inherently depth variant (DV) due to the refractive index mismatch between the imaging layers. In this study, we present a quantitative comparison among different image restoration techniques developed based on a depth-variant (DV) imaging model for fluorescence microscopy. The imaging models employed by these methods approximate DV imaging by either stratifying the object space (analogous to the discrete Fourier transform (DFT) "overlap-add" method) or image space (analogous to the DFT "overlap-save" method). We compare DV implementations based on maximum likelihood (ML) estimation and a previously developed expectation maximization algorithm to a ML conjugate gradient algorithm, using both these stratification approaches in order to assess their impact on the restoration methods. Simulations show that better restoration results are achieved with iterative methods implemented using the overlap-add method than with their implementation using the overlap-save method. However, the overlap-save method makes it possible to implement a non-iterative DV inverse filter that can trade off accuracy of the achieved result for computational speed. Results from a non-iterative regularized inverse filtering approach are also presented.

关键词： Depth-variant imaging 3D fluorescence microscopy spherical aberration image de-blurring

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Fluorescence lifetime imaging using light emitting diodes

Fluorescence lifetime imaging using light emitting diodes

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

作者： Kennedy, Gordon T. Elson, Daniel S. Munro, Ian Poher, Vincent French, Paul M. W. Neil, Mark A. A. Imperial Coll London Blackett Lab S Kensington Campus London SW7 2AZ England Imperial Coll London Dept Bioengn London SW7 2AZ England

We demonstrate fluorescence lifetime imaging using cheap, high power light emitting diodes as excitation sources. Both time domain and frequency domain fluorescence lifetime imaging techniques have been implemented at... 详细信息

ISBN: (纸本)9780819465566

关键词： fluorescence lifetime imaging LED microscopy

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Two-dimensional resolution improvement in standing wave microscopy using fast acousto-optic laser scanning

Two-dimensional resolution improvement in standing wave micr...

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

作者： Gliko, Olga Brownell, William E. Saggau, Peter Baylor Coll Med Dept Otorhinolaryngol Houston TX 77030 USA Baylor Coll Med Dept Neurosci Houston TX 77030 USA Rice Univ Dept Bioengn Houston TX USA

ISBN: (纸本)9780819474308

We present a versatile scheme for two-dimensional (2D) resolution enhancement in standing wave fluorescence microscopy (SWFM). This SWFM scheme consists of an interferometer, where both beams are focused at the back focal plane of the objective lens. Their position is controlled by a pair of acousto-optic deflectors (AODs). This results in two collimated beams that interfere in the focal plane, creating a lateral periodic excitation pattern with controlled fringe spacing and orientation. The phase of the standing wave (SW) pattern is controlled by the phase delay between two RF sinusoidal signals driving the AODs. An enlarged fluorescence image formed using the same objective lens is captured by a cooled CCD camera. Data collection involves acquiring images with excitation pattern of three equi-polar orientations (0 degrees, 60 degrees and 120 degrees) and three different phases (0 degrees, 120 degrees, 240 degrees) for each orientation. The SWFM image is algebraically reconstructed from these 9 acquired images. The SWFM image has enhanced 2D lateral resolution of about 100 nm with nearly isotropic effective point-spread function (PSF). As a result of the acousto-optic scanning, the total acquisition time can be as short as 100 mu s and is only further limited by the fluorescence intensity, as well as sensitivity and speed of the CCD camera. Utilizing acousto-optic laser scanning for advanced SWFM provides the exceptional precision and speed necessary for real-time imaging of subresolution processes in living biological systems.

关键词： super resolution fluorescence microscopy interference phase shift

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Amplitude, phase, and polarization control with a single spatial light modulator

Amplitude, phase, and polarization control with a single spa...

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

作者： Theisen, Michael J. Head, Stephen T. Brown, Thomas G. Gillmer, Steven R. Ellis, Jonathan D. Univ Rochester Inst Opt Rochester NY 14627 USA Univ Rochester Dept Mech Engn Rochester NY 14627 USA

ISBN: (纸本)9780819498625

Point spread function engineering is usually accomplished by controlling the amplitude, phase and/or polarization of the pupil fields. We analyze and test an optical design for full amplitude, phase, and polarization control of the pupil fields using a single spatial light modulator. In our scheme, the beam is spatially split into four components whose relative phases provide the four degrees of freedom necessary for amplitude, phase, and polarization control.

关键词： Spatial Light Modulators Polarization Control microscopy Pupil Engineering

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