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Mie scattering model for dual-axes confocal architecture

Mie scattering model for dual-axes confocal architecture

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

作者： Wong, Larry K. Mandella, Michael J. Holcomb, Paul Kino, Gordon S. Wang, Thomas D. Stanford Univ Sch Med Div Gastroenterol Stanford CA 94305 USA Stanford Univ Ginzton Lab Stanford CA 94305 USA Breault Res Org Inx Tucson AZ 85715 USA

ISBN: (纸本)9780819465566

Tissue scattering has a significant effect on the image resolution and light collection efficiency in confocal microscopy. The dual-axes (DA) confocal architecture has many advantages including high axial resolution with low numerical aperture lenses and long working distance for use in vivo as a microendoscope. In addition, less scattered light along the illumination path may be collected and introduced as noise. In this paper, we use Monte Carlo tissue scattering simulations to compare the dual-axes and conventional single-axis (SA) configurations. Simulation results show that the axial response for the dual axes configuration varies with pinhole size and optical thickness of scattering media in a way that differs from the single axis architecture. The DA configuration is able to filter out efficiently multiply-scattered photons and out-of-focus light, thus allowing imaging with greater tissue penetration depths to provide vertical cross-sectional images, which has significant implications for in vivo imaging.

关键词： scattering Mie theory confocal microscopy single-axis dual-axes Monte Carlo

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Multi-wavelength Digital Holographic microscopy for sub-micron topography of reflecting specimens

Multi-wavelength Digital Holographic Microscopy for sub-micr...

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

作者： Montfort, Frederic Charriere, Florian Kuehn, Jonas Colomb, Tristan Cuche, Etienne Emery, Yves 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 CHUV Ctr Neurosci Psychiat DP CH-1008 Prilly Switzerland

ISBN: (纸本)9780819465566

The study of the internal structures of specimens has a great importance in life and materials sciences. The principle of optical diffraction tomography (ODT) consists in recording the complex wave diffracted by an object, while changing the k vector of the illuminating wave. This way, the frequency domain of the specimen is scanned, allowing reconstructing the scattering potential of the sample in the spatial domain. This work presents a method for sub-micron tomographic imaging using multiple wavelengths in digital holographic microscopy. This method is based on the recording at different wavelengths equally separated in the k-domain, of the interference between an off-axis reference wave and an object wave reflected by a microscopic specimen and magnified by a microscope objective. A charged coupled device (CCD) camera records consecutively the holograms, which are then numerically reconstructed following the convolution formulation to obtain each corresponding complex object wavefronts. Their relative phases are adjusted to be equal in a given plane of interest and the resulting complex wavefronts are summed. The result of this operation is a constructive addition of complex waves in the selected plane and a destructive one in the others. Tomography is thus obtained by the attenuation of the amplitude out of the plane of interest. Numerical variation of the plane of interest enables to scan the object in depth. For the presented simulations and experiments, twenty wavelengths are used in the 480-700 nm range. The result is a sectioning of the object in slices of 725 nm thick.

关键词： digital holography topography 3D imaging

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Signal post processing in frequency domain OCT and OCM using a filter bank approach

Signal post processing in frequency domain OCT and OCM using...

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

作者： Hofer, Bernd Povazay, Boris Hermann, Boris Unterhuber, Angelika Matz, Gerald Hlawatsch, Franz Drexler, Wolfgang Cardiff Univ Dept Optometry & Vis Sci Biomed Imaging Grp Cardiff Wales Vienna Univ Technol Inst Commun & Radio Frequency Engn Vienna Austria

ISBN: (纸本)9780819465566

Current signal post processing in spectrally encoded frequency domain (FD) optical coherence microscopy (OCM) and optical coherence tomography (OCT) uses Fourier transforms in combination with non-uniform resampling strategies to map the k-space data acquired by the spectrometer to spatial domain signals which are necessary for tomogram generation. We propose to use a filter bank (FB) framework for the remapping process. With our new approach, the spectrometer is modeled as a critically sampled analysis FB, whose outputs are quantized subband signals that constitute the k-space spectroscopic data. The optimal procedure to map this data to the spatial domain is via a suitably designed synthesis FB which has low complexity. FB theory additionally states that 1) it is possible to find a synthesis FB such that the overall system has the perfect reconstruction (PR) property;2) any processing on critically sampled subband signals (as done in current schemes) results in aliasing artifacts. These perspectives are evaluated both theoretically and experimentally. We determine the analysis FB corresponding to our FD-OCM system by using a tunable laser and show that for our grating-based spectrometer - employing a CCD-line camera - the non-uniform resampling together with FFT indeed causes aliasing terms and depth dependent signal attenuation. Furthermore, we compute a finite impulse response based synthesis FB and assess the desired PR property by means of layered samples. The resulting images exhibit higher resolution and improved SNR compared to the common FFT-based approach. The potential of the proposed FB approach opens a new perspective also for other spectroscopic applications.

关键词： filter bank spectrometer spectroscopy optical coherence microscopy optical coherence tomography frequency domain tunable lasers fourier transform aliasing FFT

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Full-pupil versus divided-pupil confocal line-scanners for reflectance imaging of human skin in vivo

Full-pupil versus divided-pupil confocal line-scanners for r...

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

作者： Gareau, Dan Abeytunge, Sanjeewa Rajadhyaksha, Milind Mem Sloan Kettering Canc Ctr 1275 York Ave New York NY 10021 USA

ISBN: (纸本)9780819465566

A full-pupil confocal line-scanning microscope is under development for imaging human skin in vivo in reflectance. The new design potentially offers an alternative to current point- and line-scanners that may simplify the optics, electronics and mechanics, and lead to simpler and smaller confocal microscopes. With a combination of a cylindrical lens and an objective lens, the line-scanner creates a focused line of laser light in the object plane within tissue. An oscillating galvanometric mirror scans the focused line transverse to its axis. The backscattered light from the tissue is de-scanned and focused onto a linear CMOS detector array. Preliminary measurements of the axial line-spread function, with a 30x, 0.9-NA water immersion objective lens and illumination wavelength of 633 mn, determined the optical sectioning to be 10 pm. The new design is simple, requiring only eight optical components. However, the disadvantage is non-confocality in one dimension that results in 20% weaker sectioning than with a pointscanner, and reduced contrast in scattering tissue. The images of standard reflective targets such as a mirror and grating as well as dermis-like scattering target such as paper offer a preliminary glimpse into the performance of the line-scanner. A similar alternative design is the divided-pupil (theta) line-scanner, which provides 50% weaker sectioning than with a point scanner, but better contrast and less speckle due to the theta configuration. Such line scanners may prove useful for routine imaging of humans in clinical settings.

关键词： Confocal microscopy Skin Objectives Reflectivity In vivo imaging Mirrors Tissue optics Scanners Beam splitters Chromatic aberrations

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GPU-based visualization techniques for 3D microscopic imaging data

GPU-based visualization techniques for 3D microscopic imagin...

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conference on Computational Imaging V

作者： Wang, Qiqi Sun, Yinlong Robinson, J. Paul Purdue Univ Dept Comp Sci W Lafayette IN 47907 USA Purdue Univ Dept Biomed Engn W Lafayette IN 47907 USA

ISBN: (纸本)9780819466112

Three-dimensional (3D) microscopic imaging techniques such as confocal microscopy have become a common tool in measuring cellular structures. While computer volume visualization has advanced into a sophisticated level in medical applications, much fewer studies have been made on data acquired by the 3D microscopic imaging techniques. To optimize the visualization of such data, it is important to consider the data characteristics such as thin data volume. It is also interesting to apply the new GPU (graphics processing unit) technology to interactive volume rendering of the data. In this paper, we discuss several texture-based techniques to visualize confocal microscopy data by considering the data characteristics and with support of GPU. One simple technique generates one set of 2D textures along the axial direction of image acquisition. An improved technique uses three sets of 2D textures in the three principal directions, and creates the rendered image via a weighted sum of the images generated by blending the individual texture sets. In addition, we propose a new approach based on stencil such that textures are blended based on a stencil control. Given the viewing condition, a texel needs to be drawn only when its corresponding projection on the image plane is inside a stencil area. Finally, we have explored the use of multiple-channel datasets for flexible classification of objects. These studies are useful to optimize the visualization of 3D microscopic imaging data.

关键词： volume visualization interactive rendering GPU-based techniques texture blending stencil techniques 3D microscopic imaging confocal microscopy

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Evaluation of a new method for the determination of experimental PSF of a wide-field microscope using white-light and a linear sensor

Evaluation of a new method for the determination of experime...

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conference on Confocal, Multiphoton and Nonlinear Microscopic Imaging III

作者： Macedo, Milton P. Barata, Antonio J. Fernandes, Ana G. Correia, Carlos M. ISEC P-3030199 Coimbra Portugal Univ Coimbra CEI P-3004516 Coimbra Portugal

ISBN: (纸本)9780819467744

In the biology field there is a high demand for three-dimensional (3D) microscopy techniques owing to the need for obtaining depth information. Confocal microscopy is the reference method but it has the drawback of slow image acquisition time. Different configurations were developed to obviate this such as spinning-disk that basically consists in a set of parallel confocal microscopes and line scanning that uses line illumination so the scanning in the fast axis is not required. Amongst other 3D techniques there is the digital microscopy that normally uses a wide-field microscope to acquire 2D images (optical sections) of the specimen containing substantial contributions from out-of-focus portions. To remove this, a computational method is used that is based on the definition of a model for the process of image formation and recording. A compromise should be taken between a more accurate model with best results or a simplified one with a lower computational processing time. This work is based on a laboratory prototype of a wide-field microscope with white-light illumination using a CMOS linear image sensor. It aims at the application of one simplified model that takes advantage of a new method for the determination of experimental point spread function (PSF) that uses a specimen consisting of a set of lines in two-perpendicular directions due to the anisotropy that results from detector geometry. The 1D and 2D PSF were built. A simulation of the application of the model was developed using Matlab ((Mathworks (c)) using different simulated specimens. Its preliminary results will be presented.

关键词： PSF image formation linear sensor confocal microscopy convolution deconvolution

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Evaluation of a new method for the determination of experimental PSF of a wide-field microscope using white-light and a linear sensor

Evaluation of a new method for the determination of experime...

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Confocal, Multiphoton, and Nonlinear Microscopic Imaging III

作者： Macedo, Milton P. Barata, António J. Fernandes, Ana G. Correia, Carlos M. ISEC Instituto Superior de Engenharia de Coimbra 3030-199 Coimbra Portugal CEI Centro de Electrónica e Instrumentação Univ. Coimbra 3004-516 Coimbra Portugal

ISBN: (纸本)081946774X

In the biology field there is a high demand for three-dimensional (3D) microscopy techniques owing to the need for obtaining depth information. Confocal microscopy is the reference method but it has the drawback of slow image acquisition time. Different configurations were developed to obviate this such as spinning-disk that basically consists in a set of parallel confocal microscopes and line scanning that uses line illumination so the scanning in the fast axis is not required. Amongst other 3D techniques there is the digital microscopy that normally uses a wide-field microscope to acquire 2D images (optical sections) of the specimen containing substantial contributions from out-of-focus portions. To remove this, a computational method is used that is based on the definition of a model for the process of image formation and recording. A compromise should be taken between a more accurate model with best results or a simplified one with a lower computational processing time. This work is based on a laboratory prototype of a wide-field microscope with white-light illumination using a CMOS linear image sensor. It aims at the application of one simplified model that takes advantage of a new method for the determination of experimental point spread function (PSF) that uses a specimen consisting of a set of lines in two-perpendicular directions due to the anisotropy that results from detector geometry. The ID and 2D PSF were built. A simulation of the application of the model was developed using Matlab ((Mathworks ©) using different simulated specimens. Its preliminary results will be presented. © 2007 SPIE-OSA.

关键词： image processing

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image enhancement for digital visualization of three dimensional integral imaging

Image enhancement for digital visualization of three dimensi...

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

作者： Ponce-Diaz, Rodrigo Alvarez-Elizondo, Martha Saenz, Julia Javidi, Bahram Inst Tecnol & Estudios Super Monterrey Av Eugenio Garza Sada 2501CP 64849 Monterrey NL Mexico Univ Connecticut Storrs CT 06269 USA

ISBN: (纸本)0819461326

In this work, we are proposing the use of digital image enhancement in three dimensional (3D) Integral Imaging (InI) applying in small objects visualization. We used a well known image filter algorithm for enhance the edges and detail information of the 3D reconstruction InI image via unsharp masking. Small objects as bugs were recorded in an elemental image array, image processed and digital reconstructed as 3D objects. We implemented the algorithm over the elemental image array as usual and using an innovative technique that involves a simple digital reconstruction algorithm by quadruple pixel extraction. Digital results show an improvement in details visualization, which have potential application in 3D microscopy.

关键词： integral imaging image enhancement three dimensional processing

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High-throughput hyperspectral microscopy

High-throughput hyperspectral microscopy

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

作者： Gehm, M. E. Brady, D. J. Duke Univ Fitzpatrick Ctr Photon Durham NC 27708 USA

ISBN: (纸本)0819461326

A hyperspectral imager provides a 3-D data cube in which the spatial information (2-D) of the image is complemented by spectral information (1-D) about each spatial location. A static, high-throughput spectrometer design previously developed by our group can be used as the spectral engine in a high-throughput hyperspectral imager that avoids the Fourier undersampling issues present in previous dispersive designs. We present the theory for both pushbroom and tomographic operation and describe experimental results from our proof-of-concept implementation of a hyperspectral microscope.

关键词： hyperspectral imaging multidimensional microscopy

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Adaptive optics for three-dimensional optical data storage and micromachining

Adaptive optics for three-dimensional optical data storage a...

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

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

ISBN: (纸本)0819461326

Many high resolution optical methods are affected by the presence of optical aberrations. These include microscopy, three-dimensional optical data storage and optical micromachining. We investigate the use of adaptive aberration correction applied to these techniques. In particular, it is shown how a deformable membrane mirror can be used to correct the aberrations when focusing deep into a multilayer optical data storage medium for both recording and read-out of data. Aberration correction for optical micromaching deep inside a substrate is also demonstrated.

关键词： Aberration correction Optical storage Monochromatic aberrations Adaptive optics Micromachining Microscopes 3D optical data storage Confocal microscopy Data storage Objectives

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