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检索条件"任意字段=Conference on 3-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing IX"

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High resolution optical projection tomographic microscopy for 3D tissue imaging

High resolution optical projection tomographic microscopy fo...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XVIII

作者： Miao, Qin Hayenga, Jon Meyer, Michael G. Neumann, Thomas Patten, Florence Nelson, Alan C. Seibel, Eric J. Univ Washington Dept Bioengn Seattle WA 98195 USA VisionGate Inc Gig Harbor WA 98335 USA Univ Washington Dept Mech Engn Seattle WA 98195 USA

ISBN: (纸本)9780819484413

Optical projection tomography (OPT) requires a large depth of field (DOF) of a low numerical aperture (NA) lens resulting in low resolution. However, DOF of a high NA objective can be extended by scanning the focal plane through the sample. This extended DOF image is called pseudoprojection, which is used by optical projection tomographic microscope (OPTM) for tomographic reconstruction. The advantage of OPTM is the acquisition of relatively high resolution and large depth of field concurrently. This method requires the working distance of the lens to be larger than the size of the sample, so proper lens should be chosen for samples of different sizes. In this paper, we imaged hematoxylin stained muntjac cells inside capillary tube with two different sizes. Two objective lenses with different NA are used for these two tubes. Experimental results show that resolution improves over 10 times in OPTM compared to conventional OPT, which make it possible for OPTM technique to resolve sub-cellular features for large samples. Therefore, OPTM can be used for 3D histological analysis of hematoxylin & eosin (H&E) stained biopsy specimen with sub-cellular resolution in the future.

关键词： Three-dimensional microscopy Tomographic imaging Tissue imaging Needle biopsy

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Further developments in addressing depth-variant 3D fluorescence microscopy imaging

Further developments in addressing depth-variant 3D fluoresc...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXI

作者： Ghosh, Sreya Schaefer, Lutz Schuster, Dietwald Preza, Chrysanthe Univ Memphis Dept Elect & Comp Engn Memphis TN 38152 USA Adv Imaging Methodol Consultat Kitchener ON Canada Regensburg Univ Appl Sci Regensburg Germany

ISBN: (纸本)9780819498625

Three-dimensional (3D) imaging with optical sectioning microscopy uses computational methods to obtain the true fluorescence distribution by ameliorating the effect of defocus, spherical aberration and noise. Inverse algorithms improve image quality at a fraction of the cost of implementing an optical system by accurate modeling of the imaging system. Good inverse imaging algorithms need to be accurate as well as fast. Better understanding of the image formation model is vital to obtain improved restoration through model-based algorithms. Forward imaging models based on a depth-varying point-spread function (DV-PSF) leads to a substantial improvement in the resulting images because it accounts for depth-induced aberrations present in the imaging system. PSFs at every layer can be represented using their principal components. Computation of the forward imaging model using a principal component analysis (PCA) representation of the DV-PSF requires fewer convolutions than a strata based approach investigated in the past. In this paper we present a new algorithm for maximum likelihood image restoration developed based on a PCA representation of the DV-PSF and an accelerated conjugate gradient (CG) iteration scheme. Results obtained with this PCA-CG algorithm from both simulated and experimental fluorescence microscope data are discussed and compared with results obtained from a CG iteration method based on the strata model and linear interpolation of the DV-PSF. The performance of the PCA-CG algorithms is shown to be promising for practical applications.

关键词： Point spread functions Principal component analysis 3D image processing Expectation maximization algorithms 3D modeling image restoration Algorithm development Computer simulations Luminescence Reconstruction algorithms

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Dual-detection confocal microscopy: high-speed surface profiling without depth scanning 13

Dual-detection confocal microscopy: high-speed surface profi...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXIII

作者： Lee, Dong-Ryoung Gweon, Dae-Gab Yoo, Hongki Korea Adv Inst Sci & Technol KAIST Dept Mech Engn Nano Optomechatron Lab 291 Daehak Ro Daejeon 34141 South Korea Hanyang Univ Dept Biomed Engn Biomed Opt & Photomed Lab 222 Wangsimni Ro Seoul 04763 South Korea

ISBN: (纸本)9781628419474

We propose a new method for three-dimensional (3-D) imaging without depth scanning that we refer to as the dual-detection confocal microscopy (DDCM). Compared to conventional confocal microscopy, DDCM utilizes two pinholes of different sizes. DDCM generates two axial response curves which have different stiffness according to the pinhole diameters. The two axial response curves can draw the characteristics curve of the system which shows the relationship between the axial position of the sample and the intensity ratio. Utilizing the characteristic curve, the DDCM reconstructs a 3-D surface profile with a single 2-D scanning. The height of each pixel is calculated by the intensity ratio of the pixel and the intensity ratio curve. Since the height information can be obtained directly from the characteristic curve without depth scanning, a major advantage of DDCM over the conventional confocal microscopy is a speed. The 3-D surface profiling time is dramatically reduced. Furthermore, DDCM can measure 3-D images without the influence of the sample condition since the intensity ratio is independent of the quantum yield and reflectance. We present two types of DDCM, such as a fluorescence microscopy and a reflectance microscopy. In addition, we extend the measurement range axially by varying the pupil function. Here, we demonstrate the working principle of DDCM and the feasibility of the proposed methods.

关键词： Confocal microscopy three-dimensional microscopy high speed imaging

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Spectral domain phase microscopy: A new tool for measuring cellular dynamics and cytoplasmic flow

Spectral domain phase microscopy: A new tool for measuring c...

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

作者： McDowell, EJ Choma, MA Ellerbee, AK Izatt, JA Duke Univ Dept Biomed Engn Durham NC 27708 USA

ISBN: (纸本)0819456756

Broadband interferometry is an attractive technique for the detection of cellular motions because it provides depth-resolved interferometric phase information via coherence gating. Here a phase sensitive technique called spectral domain phase microscopy (SDPM) is presented. SDPM is a functional extension of spectral domain optical coherence tomography that allows for the detection of cellular motions and dynamics with nanometer-scale sensitivity. This sensitivity is made possible by the inherent phase stability of spectral domain OCT combined with common-path interferometry. The theory that underlies this technique is presented, the sensitivity of the technique is demonstrated by the measurement of the thermal expansion coefficient of borosilicate glass, and the response of an Amoeba proteus to puncture of its cell membrane is measured. We also exploit the phase stability of SDPM to perform Doppler flow imaging of cytoplasmic streaming in A. proteus. We show reversal of cytoplasmic flow in response to stimuli, and we show that the cytoplasmic flow is laminar (i.e. parabolic) in nature. We are currently investigating the use of SDPM in a variety of different cell types.

关键词： Optical Coherence Tomography interferometry interference microscopy Doppler flow imaging cellular dynamics

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Projection multiplexing for enhanced acquisition speed in holographic tomography 26

Projection multiplexing for enhanced acquisition speed in ho...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XXVI

作者： Kus, Arkadiusz Baczewska, Maria Ziemczonok, Michal Kujawinska, Malgorzata Warsaw Univ Technol Inst Micromech & Photon Sw Boboli 8 St PL-02525 Warsaw Poland

ISBN: (纸本)9781510624092

In holographic tomography (HT), the 3D refractive index distribution within weakly-scattering, phase-only biological object is retrieved. This key property of the technique is one of its most significant strengths compared to labelling-based methods of cell analysis such as fluorescence microscopy. As a consequence, however, it is required to acquire a set of holograms at several viewing directions, which hinders the measurement speed. In this paper we explore the prospect of multiplexing projections in order to decrease the number of scanning positions required for the full measurements. The presented analysis is based on experimental data acquired in a limited-angle holographic tomography system and emulates the performance of a spatial-light-modulator-based system in which multiple projections may be acquired simultaneously by generating a distribution of multiple point sources in the Fourier plane of the condenser lens. For this reason, the increase in acquisition speed strictly depends on the number of multiplexed holograms and results in decreased reconstruction quality. The performance of the system is demonstrated and analyzed with biological objects - human keratinocyte cells.

关键词： Holographic tomography optical diffraction tomography tomographic phase microscopy quantitative phase imaging 3D refractive index hologram multiplexing

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Absolute and relative quantification and calibration for sectioning fluorescence microscopy using standardized uniform fluorescent layers and SIPchart based correction procedures

Absolute and relative quantification and calibration for sec...

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conference on Three-dimensional and multidimensional microscopy - image acquisition and processing XIV

作者： Zwier, J. M. Oomen, L. Brocks, L. Jalink, K. Brakenhoff, G. J. Univ Amsterdam Swammerdam Inst Life Sci Kruislaan 316 NL-1098 SM Amsterdam Netherlands Netherlands Canc Inst NL-1066 CX Amsterdam Netherlands

ISBN: (纸本)9780819465566

The total or integrated fluorescence intensity of a through-focus series of a thin standardized uniform fluorescent or calibration layer is shown to be suitable for image intensity correction and calibration in sectioning microscopy. This integrated intensity can be derived from the earlier introduced Sectioned Imaging Property or SIPcharts,(1) derived from the 3D layer datasets. By correcting the 3D image of an object with the 3D image of the standardized uniform fluorescent layer obtained under identical conditions one is able to express the object fluorescence in units fluorescence of the calibration layer. With object fluorescence intensities in 'fluorescence layer unit's or FLU's the object image intensities becomes independent of microscope system and imaging conditions. A direct result is that the often-appreciable lateral intensity variations present in confocal microscopy are eliminated (shading correction). Of more general value is that images obtained with different objectives, magnifications or from different microscope systems can be quantitatively related to each other. The effectiveness of shading correction and relating images obtained under various microscope conditions is demonstrated on images of standard fluorocent beads. Expressing the object fluorescence in FLU units seems to be a promising approach for general quantification of sectioning imaging enabling cross-correlation of imaging results over time and between imaging systems.

关键词： fluorescence microscopy shading correction sectioning microscopy confocal microscopy sectioned imaging 3D microscopy image correction SIPcharts

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Super-resolution stereo light microscopy for real-time tasks using normal flow and geometric constraints

Super-resolution stereo light microscopy for real-time tasks...

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

作者： Danuser, G Swiss Fed Inst Technol ETH BioMicroMetr Grp Lab Biomech CH-8952 Schlieren Switzerland

ISBN: (纸本)081943535X

Prior knowledge about the observed scene represents the key to recovering frequencies beyond the passband of an imaging system (super-resolution). With regard to microscopy mainly two super-resolution mechanisms have been reported: 1.) analytic continuation of the frequency spectrum and 2.) constrained image deconvolution. This paper describes an alternative super-resolution mechanism. Prior knowledge is introduced on a higher, more symbolic level of visual inference. We exemplify our concept based on the 3D reconstruction of a micro-pipette moving in close proximity to immobile target objects. Information about the shape and the mobility of the pipette is incorporated in order to localize the pipette tip at sub-Rayleigh distances to the target. The algorithm was tested in a micro-robot environment. A machine vision module using stereo light microscopy automatically controlled the manipulation of microscopic objects, e.g. latex beads or diamond mono-crystals. In the theoretical part of the paper we prove that knowledge of the form "the pipette has moved between two consecutive frames of the movie" must result in a twofold increase of resolution. We used the normal flow of an image to decode positional measures from motion. In practice, super-resolution factors between 3 to 5 were obtained. The additional gain originates from the geometric constraints which were included to reconstruct the pipette position.

关键词： light microscopy stereo super-resolution machine vision normal flow geometric constraints

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Study of the spatial point spread function with depth in two-photon microscopy

Study of the spatial point spread function with depth in two...

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conference on 3-dimensional and multidimensional microscopy - image acquisition and processing VI

作者： Dunn, A Wallace, V Coleno, M So, P Tromberg, B Univ Calif Irvine Beckman Laser Inst & Med Clin LAMMP Irvine CA 92612 USA

ISBN: (纸本)0819430757

The spatial point spread function is studied for two-photon fluorescence imaging in turbid media. Using a Monte Carlo based model the effects of tissue optical properties on the point spread function, fluorescence generation, and fluorescence collection are studied. The simulated results are compared to measured point spread functions at different depths within a scattering sample. Results indicate that the limiting factor in two-photon imaging is a loss of signal caused by scattering rather than a loss of resolution.

关键词： two-photon fluorescence microscopy Monte Carlo

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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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The spectral topography of histopathology samples

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

作者： Lerner, JM Lu, TW Vari, S LightForm Inc Belle Mead NJ 08502 USA

ISBN: (纸本)0819427004

The goal of imaging spectroscopy is to obtain independent spectra from individual objects in a field-of-view. In the case of biological materials, such as histopathology samples, it has been well established that spectral characteristic can be indicative of specific diseases including cancer. Diagnosis can be enhanced by the use of probes and stains to indicate the presence of individual genome or other biologically active cell components or substances. To assess a specimen through a microscope is directly analogous to observing the Earth from space to assess natural features. This paper describes a simple and inexpensive Imaging Spectrometer, with an origin in remote sensing, that demonstrates that it is possible to rapidly identify evidence of disease in histopathology samples using spatially resolved spectral data. The PARISS imaging spectrometer enables a researcher to acquire multispectral images that yield functional maps, showing what and where biological molecules are located within a structure. It is the powerful combination of imaging and spectroscopy that provides the tools not readily available to the Life Sciences. The PARISS system incorporates a powerful hybrid Neural Network (HNN) analysis to break the data logjam that is often associated with the acquisition and processing of multiple spectra.

关键词： imaging spectroscopy spectral topography histopathology spectroscopy micro-imaging microscopy

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