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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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Biological cell (pollen grain) refractive index tomography with digital holographic microscopy

Biological cell (pollen grain) refractive index tomography w...

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

作者： Charriere, Florian Cuche, Etienne Marquet, Pierre Depeursinge, Christian Ecole Polytech Fed Lausanne Imaging & Appl Opt Inst CH-1015 Lausanne Switzerland Lyncee Tec SA PSE A CH-1015 Lausanne Switzerland CHU Vaudois Ctr Neurosci Psychiat Dept Psychiat CH-1008 Lausanne Switzerland

ISBN: (纸本)0819461326

This paper present a novel approach to perform the tomography of biological specimen based on Digital Holographic microscopy (DHM). A hologram results from the interference between a reference wave and an object wave reflected from or transmitted through a sample. In the hologram, both amplitude and phase of the field transmitted through the object are registered. In DHM, the object field is recovered when the hologram is processed by a digitally computed replica of the reference wave, allowing quantitative measurement of both phase and amplitude. Phase measurements provide high accuracy optical path length measurements across the specimen along the optical axis. To proceed to a tomographic reconstruction of the refractive index of the sample based on this quantitative phase measurement, such 2-dimentionnal data must be recorded for different sample orientations covering an angle of 180 degrees to cover all the object spatial frequencies in the reciprocal space. The representation of the data in function of the angle is known as a sinogram. The 3-dimentionnal refractive index can then be reconstructed from the sinograms by a filtered backprojection algorithm. In our system, the specimen is inserted in a glass micropipette to permit its rotation. To our knowledge, a quantitative tomography of the refractive index of a pollen cell with a resolution in the micron range is presented for the first time.

关键词： holography microscopy phase-contrast tomography refractive index cell imaging

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Multiphoton imaging of basal cell carcinoma (BCC)

Multiphoton imaging of basal cell carcinoma (BCC)

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

作者： Cicchi, R. Carli, P. Massi, D. Sestini, S. Stambouli, D. Pavone, F. S. Univ Florence Dept Dermatol Pergola 58 I-50121 Florence Italy

ISBN: (纸本)0819461326

We used two-photon microscopy towards the imaging of cutaneous basal cell carcinoma (BCC). Our aim was to evaluate the morphology of BCC using two-photon fluorescence excitation and to establish a correlation with histopathology. We built a custom two-photon microscope and we measured the system capabilities. The system allowed to perform a preliminary measurement on a fresh human skin tissue sample. A human skin tissue sample of BCC excised during dermatological surgery procedures were used. The clinical diagnosis of BCC was confirmed by subsequent histopathological examination. The sample was imaged using endogenous tissue fluorescence within 2-3 hours from the excision with a two photon laser scanning fluorescence microscope. The acquired images allowed an obvious discrimination of the neoplastic areas toward normal tissue, based on morphological differences and aberrations of the intensity of the fluorescence signal. Our results showed that BCC tissue has a more homogeneous structure in comparison to normal tissue as well as a higher fluorescent response. The images obtained by two photon microscopy were further compared to the images acquired by an optical microscope after the conventional histopathological examination on one part of the respective sample. Our suggested method may represent a new diagnostic tool that improves the diagnostic accuracy of clinical examination alone, enabling the accurate discrimination of basal cell carcinoma from normal tissue.

关键词： multiphoton microscopy basal cell carcinoma diagnosis optical biopsy

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A novel stochastic resolution criterion for fluorescence microscopes

A novel stochastic resolution criterion for fluorescence mic...

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

作者： Ram, Sripad Ward, E. Sally Ober, Raimund J. Univ Texas Southwestern Med Ctr Dallas Ctr Immunol NB9-1066000 Harry Hines Blvd Dallas TX 75235 USA Univ Texas Southwestern Med Ctr Dallas Univ Texas Arlington Joint Biomed Engn Grad Program Dallas TX 75235 USA Univ Texas Dallas Dept Elect Engn Richardson TX 75083 USA

ISBN: (纸本)0819461326

Rayleigh's criterion is widely in optical microscopy to determine the resolution of microscopes. Despite its widespread use, it is well known that this criterion is heuristic and does not consider the actual measurement process. For instance, it has been pointed out that the resolution of a fluorescence microscope can, in principle, exceed Rayleigh's criterion when distance determination between two point sources is postulated as a parameter estimation problem. In fact, recent results from single molecule fluorescence experiments show that the location of two closely spaced point sources with distance of separation well below Rayleigh's criterion can be accurately estimated. These results suggest that Rayleigh's criterion is inadequate for current microscope techniques. Here, by adopting an information-theoretic stochastic framework, we re-visit the resolution problem and derive a new stochastic resolution criterion that provides a limit to the accuracy with which the distance between two point sources can be determined. Our results predict that the distance between two point sources can be determined to an arbitrary accuracy provided a sufficient number of photons are detected. The new criterion is given in terms of quantities such as the expected number of detected photons, the numerical aperture of the objective lens and the wavelength of the detected photons. We also investigate how the new resolution measure is influenced by deteriorating experimental factors such as pixelation of the detector and additive noise sources.

关键词： Photons Molecules image resolution Stochastic processes Microscopes Sensors Luminescence Data acquisition microscopy Objectives

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A study of Gaussian approximations of fluorescence microscopy PSF models

A study of Gaussian approximations of fluorescence microscop...

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

作者： Zhang, Bo Zerubia, Josiane Olivo-Marin, Jean-Christophe Inst Pasteur Unite Analyse Images Quantitat 25-28 Rue Dr Roux F-75015 Paris France Ecole Natl Super Telecommun Bretagne F-75013 Paris France Inst Natl Rech Informat & Automat F-06902 Sophia Antipolis France

ISBN: (纸本)0819461326

Despite the availability of rigorous physical models of microscopy point spread functions (PSFs), approximative PSFs, particularly separable Gaussian approximations are widely used in practical microscopic data processing. In fact, compared with a physical PSF model, which usually involves non-trivial terms such as integrals and infinite series, a Gaussian function has the advantage that it is much simpler and can be computed much faster. Moreover, due to its special analytical form, a Gaussian PSF is often preferred to facilitate the analysis of theoretical models such as Fluorescence Recovery After Photobleaching (FRAP) process and of processing algorithms such as EM deconvolution. However, in these works, the selection of Gaussian parameters and the approximation accuracy were rarely investigated. In this paper, we present a comprehensive study of Gaussian approximations for diffraction-limited 2D/3D paraxial/non-paraxial PSFs of Wide Field Fluorescence microscopy (WFFM), Laser Scanning Confocal microscopy (LSCM) and Disk Scanning Confocal microscopy (DSCM) described using the Debye integral. Besides providing an optimal Gaussian parameter for the 2D paraxial WFFM PSF case, we further derive nearly optimal parameters in explicit forms for each of the other cases, based on Maclaurin series matching. Numerical results show that the accuracy of the 2D approximations is very high (Relative Squared Error (RSE) < 2% in WFFM, < 0.3% in LSCM and < 4% in DSCM). For the 3D PSFs, the approximations are average in WFFM (RSE similar or equal to 16 - 20%), accurate in DSCM (RSE similar or equal to 3 - 6%) and nearly perfect in LSCM (RSE similar or equal to 0.3 - 0.5%).

关键词： Gaussian approximation of PSF Wide Field Fluorescence microscopy Laser Scanning Confocal microscopy Disk Scanning Confocal microscopy microscopic data processing

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Optimum conditions for high-quality 3D reconstruction in confocal scanning microscopy

Optimum conditions for high-quality 3D reconstruction in con...

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

作者： Kim, Taehoon Kim, Taejoong Lee, SeungWoo Gweon, Dae-Gab Seo, Jungwoo Korea Adv Inst Sci & Technol Dept Engn Mech Nano Opto Mechatron Lab Guseong Dong 373-1 Taejon 305701 South Korea Pusan Natl Univ Dept Nanomech Engn Busan South Korea

ISBN: (纸本)0819461326

Confocal Scanning microscopy (CSM) is very useful to reconstruct 3D image of Bio-cells and the objects that have specification shape in higher axial and lateral resolution and widely used as measurement instrument. A 3D reconstruction is used to visualize confocal images and consists of following processes. The First process is to get 3D data by collecting a series of images at regular focus intervals (Optical Sectioning). The Second process is to fit a curve to a series of 3D data points each pixel. The Third process is to search height information that has maximum value from curve-fitting. However, because of various systematic errors (NOISE) occurred when collecting the information of images through Optical Sectioning and large peak deviation occurred from curve-fitting error, high quality 3D reconstruction is not expected. Also, it takes much time to 3d Reconstruction by using many 3D data in order to acquire high quality and much cost to improve signal-to-noise (SNR) using a higher power laser. So, we are going to define SNR, peak deviation and the order of curve-fitting as important factors and simulate the relation between the factors in order to find a optimum condition for high quality 3D reconstruction in Confoal Scanning microscopy. If we use optimum condition obtained by this simulation, using a suitable SNR and the suitable number of data and the suitable nth order curve-fitting, small peak deviation is expected and then, 3D reconstruction of little better quality is expected. Also, it is expected to save.

关键词： Confocal Scanning microscopy Optical Sectioning high quality 3D reconstruction SNR NOISE optimum conditions

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Proceedings of SPIE: three-dimensional and multidimensional microscopy: image acquisition and processing ix

Proceedings of SPIE: Three-dimensional and Multidimensional ...

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three-dimensional and multidimensional microscopy: image acquisition and processing ix

This Volume 4621 of the conference proceedings contains 16 papers. Topics discussed include three dimensional and multidimensional microscopy, new methods and instruments, microscope characterization and assessment an... 详细信息

关键词： Microscopic examination

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Progress in Biomedical Optics and Imaging - three-dimensional and multidimensional microscopy: image acquisition and processing XII

Progress in Biomedical Optics and Imaging - Three-Dimensiona...

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three- dimensional and Multidomensional microscopy: image acquisition and processing XII

The proceedings contain 23 papers from the conference on Progress in Biomedical Optics and Imaging - three-dimensional and multidimensional microscopy: image acquisition and processing XII. The topics discussed include: using out-of-focus light to improve image acquisition time in confocal microscopy;three-dimensional optical computed-tomography microscope;image estimation for structured-illumination microscopy;microlymphatic flow using fast video microscopy;confocal reflectance theta line-scanner for imaging tissues in vivo;multimodal optical microscope for imaging biological systems;spectral domain phase microscopy: a new tool for measuring cellular dynamics and cytoplasmic flow;and an algorithm for image stitching and blending.

关键词： Imaging systems

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three-dimensional multiple nanometric local probes microscopy

Three-dimensional multiple nanometric local probes microscop...

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

作者： Moneron, G Dubois, A Boccara, AC CNRS UPA A0005 Ecole Super Phys Chim Ind Lab Opt Phys F-75231 Paris France

ISBN: (纸本)0819456756

We present a microscope set-up designed to produce three-dimensional images of the internal structures of various samples with high spatial resolution (a few tens of nanometers in axial and transverse directions). This level of resolution is reached by the use of nanometric sub-wavelength spherical gold beads as multiple local probes, dispersed in the hollow structures. The exploration, by Brownian motion, of the internal structures allows their three-dimensional reconstruction.

关键词： interference microscopy three-dimensional microscopy optical coherence tomography white-light interferometry scanning probe microscopy

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image estimation for structured-illumination microscopy

Image estimation for structured-illumination microscopy

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

作者： Conchello, JA Oklahoma Med Res Fdn Program Mol Cell & Dev Biol Oklahoma City OK 73104 USA

ISBN: (纸本)0819456756

When recording three-dimensional (3D) images by the method of optical sectioning microscopy, each optical section contains the in-focus information plus out-of-focus contributions that obscure the in-focus detail and reduce contrast. There are several methods to remove or prevent the out-of-focus contributions from the stack of optical sections. One such method is image estimation -the use of a computer program based on a mathematical description of the microscope to remove the out-of-focus contributions. Another method is the use of structured illumination and a simple arithmetic operation to obtain a image that in which the out-of-focus contributions are greatly reduced. We derived a method for image estimation that uses the images collected from the structured-illumination microscope. The method improves the resolution of small detail over that possible with the structured illumination using the simple arithmetic formula.

关键词： image analysis Expectation maximization algorithms Microscopes microscopy 3D image processing Confocal microscopy image acquisition image processing Point spread functions Luminescence

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Scanning microscopy with extended depth of focus

Scanning microscopy with extended depth of focus

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

作者： Juskaitis, R Botcherby, EJ Wilson, T Univ Oxford Dept Engn Sci Oxford England

We demonstrate a simple and light-efficient way of generating non-diffracting Bessel beams for use in confocal microscopy. A number of imaging modalities using such beams is discussed. Preliminary experimental results... 详细信息

ISBN: (纸本)0819456756

关键词： confocal extended depth of focus Bessel beam fluorescence microscopy polarisation multi-photon

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