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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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Cell morphology classification in phase contrast microscopy image reducing halo artifact

Cell morphology classification in phase contrast microscopy ...

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

作者： Kang, Mi-Sun Song, Soo-Min Lee, Hana Kim, Myoung-Hee Ewha Womans Univ Dept Comp Sci & Engn Seoul 120750 South Korea

ISBN: (纸本)9780819488701

Since the morphology of tumor cells is a good indicator of their invasiveness, we used time-lapse phase-contrast microscopy to examine the morphology of tumor cells. This technique enables long-term observation of the activity of live cells without photobleaching and phototoxicity which is common in other fluorescence-labeled microscopy. However, it does have certain drawbacks in terms of imaging. Therefore, we first corrected for non-uniform illumination artifacts and then we use intensity distribution information to detect cell boundary. In phase contrast microscopy image, cell is normally appeared as dark region surrounded by bright halo ring. Due to halo artifact is minimal around the cell body and has non-symmetric diffusion pattern, we calculate cross sectional plane which intersects center of each cell and orthogonal to first principal axis. Then, we extract dark cell region by analyzing intensity profile curve considering local bright peak as halo area. Finally, we examined cell morphology to classify tumor cells as malignant and benign.

关键词： cell morphology phase contrast microscopy image halo artifact cell classification

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LABEL-FREE CLASSIFICATION OF CELL TYPES BY IMAGING OF CELL MEMBRANE FLUCTUATIONS USING LOW-COHERENT FULL-FIELD QUANTITATIVE PHASE microscopy

LABEL-FREE CLASSIFICATION OF CELL TYPES BY IMAGING OF CELL M...

引用

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

作者： Yamauchi, Toyohiko Sugiyama, Norikazu Sakurai, Takashi Iwai, Hidenao Yamashita, Yutaka Hamamatsu Photon KK Hamakita Ku 5000 Hirakuchi Hamamatsu Shizuoka 4348601 Japan MIT Spectroscopy Lab Cambridge MA 02139 USA Hamamatsu Univ Sch Med Photon Med Res Ctr Hamamatsu Shizuoka 4313192 Japan

ISBN: (纸本)9780819479662

Cell membrane motions of living cells are quantitatively measured in nanometer resolution by low-coherent full-field quantitative phase microscopy. Our setup is based on a full-field phase shifting interference microscope with a very low-coherent light source. The reflection mode configuration and the low-coherent illumination make it possible to differentiate the weak reflection light from the cell membrane from the strong reflection from the glass substrate. Two cell populations are quantitatively assessed by the power spectral density of the cell surface motion and show different trends.

关键词： quantitative phase microscope cell imaging plasma membrane

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Supercritical self-interference fluorescence microscopy for full-field membrane imaging

Supercritical self-interference fluorescence microscopy for ...

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

作者： Barroca, Thomas Bon, Pierre Leveque-Fort, Sandrine Fort, Emmanuel ESPCI ParisTech Inst Langevin Ctr Imageries Plasmon Appl CNRSUMR 7587 1 Rue Jussieu F-75231 Paris 05 France Univ Paris Sud CNRS CLUPS UMR 8214 F-91405 Orsay France

ISBN: (纸本)9780819493583

We present a new technique based on the self-interference of Supercritical Angle Fluorescence (SAF) emission in order to perform full-field cell membrane imaging. We show that our Point Spread Function (PSF) engineering technique allows us to obtain a 100 nm axial sectioning while conserving the original lateral resolution of the microscope. The images are acquired using an optical module that can be connected to any fluorescent microscope to simultaneously monitor in real time both the cell membrane and in-depth phenomena.

关键词： Cell membrane imaging Supercritical Angle Fluorescence (SAF) Fluorescence microscopy Dictyostelium discoideum

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Fabrication and characterization of novel microsphere-embedded optical devices for enhancing microscopy resolution 25

Fabrication and characterization of novel microsphere-embedd...

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conference on Three-dimensional and multidimensional microscopy: image acquisition and processing XXV

作者： Darafsheh, Arash Washington Univ Sch Med Dept Radiat Oncol St Louis MO 63110 USA

ISBN: (数字)9781510614840

ISBN: (纸本)9781510614840;9781510614833

Microsphere-assisted imaging can be incorporated onto conventional light microscopes allowing wide-field and flourescence imaging with enhanced resolution. We demonstrated that imaging of specimens containing sub diffraction-limited features is achievable through high-index microspheres embedded in a transparent thin film placed over the specimen. We fabricated novel microsphere-embedded microscope slides composed of barium titanate glass microspheres (with diameter 10-100 lam and refractive index-4.9-2.2) embedded in a transparent polydimethylsiloxane (PDMS) elastomer layer with controllable thickness. We characterized the imaging performance of such microsphere-embedded devices in white-light microscopies, by measuring the imaging resolution, field-of-view, and magnification as a function of microsphere size. Our results inform on the design of novel optical devices, such as microsphere-embedded microscope slides for imaging applications.

关键词： Super-resolution microscopy nanoscopy microsphere barium titanate PDMS

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microscopy using micro-pixellated light emitting diodes

Microscopy using micro-pixellated light emitting diodes

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

作者： Poher, V. Kennedy, G. T. Elson, D. S. French, P. M. W. Neil, A. A. A. Imperial Coll London Blackett Lab South Kensington Campus London SW7 2AZ England

ISBN: (纸本)9780819465566

In this paper we demonstrate structured illumination microscopy, a cheap and flexible method of obtaining optical sectioning in wide-field microscopy, and we investigate line-scanning microscopy, both using the same microelement LED. In our first experiment we demonstrate structured illumination using a custom-designed microelement LED consisting of 120 individually addressable stripes emitting at 470 mn. An electrical driver was designed to produce a programmable grid pattern and the device was located in an Olympus BX41 fluorescence microscope in critical illumination configuration. By using an integrated solid-state alternative to a conventional piezo-actuated grid and separate illumination source, we improved the speed and accuracy of the system, reducing the artefacts due to the errors in the grid positions. Additionally, we investigated the use of the same LED device as a scanning source for confocal line-scanning microscopy. After each line scanning, an image was acquired using a CCD camera and the out-offocus light was rejected by a post-processing method.

关键词： structured illumination microscopy line scanning confocal microscopy micro-stripe light emitting diode

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Time-Resolved Single Molecule microscopy coupled with Atomic Force microscopy

Time-Resolved Single Molecule Microscopy coupled with Atomic...

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

作者： Koenig, M. Koberling, F. Schulz, O. Ros, R. Fore, S. Sackrow, M. Trautmann, S. Erdmann, R. PicoQuant GmbH Rudower Chaussee 29 D-12489 Berlin Germany Arizona State Univ Dept Phys Tempe AZ 85287 USA PicoQuant Photon North Amer Inc Westfield MA 01085 USA

ISBN: (纸本)9780819488701

Time-resolved confocal microscopy is well established to image spectral and spatial properties of samples in biology and material science. Atomic Force microscopy (AFM) in addition enables to investigate properties which are not optically addressable or are hidden by the diffraction limited optical resolution. We present a straight forward combination of single molecule sensitive time-resolved confocal microscopy with different commercially available AFMs. Besides an extra of information about for example a cell surface, the AFM tip can also be used to manipulate the sample on a nanometer scale down to the single molecule level.

关键词： Confocal microscopy Atomic Force microscopy AFM quenching time-resolved fluorescence sub diffraction single molecule

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A linear algorithm for quantitative measure of corneal collagen fiber orientation using second harmonic generation microscopy 13

A linear algorithm for quantitative measure of corneal colla...

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

作者： McLean, James DiMarzio, Charles Northeastern Univ 360 Huntington Ave Boston MA 02115 USA

ISBN: (纸本)9781628419474

There is significant interest in the scientific community to develop a reliable and precise quantification of the direction of collagen fibers within the cornea in order to learn more about how collagen contributes to the cornea's shape and structure. Previous work has shown that quantification of these fibers' orientations is possible using Second Harmonic Generation (SHG) microscopy, a modality that utilizes the non-centrosymmetric properties of collagen to obtain details of their macromolecular structure. Many attempts at using SHG to this purpose result in whole volume approximations which do not take into account variations through the depth of the cornea. Additionaly, other algorithms have used non-linear processes which limit computation time and result in sensitivity to sample area, resolution, and illumination. We propose a linear method for quantification of collagen fiber orientation which utilizes the precise sectioning properties of SHG and is independent of non-linear artifacts to aid in further classification of cornea structure.

关键词： Second Harmonic Generation collagen cornea orientation quantification

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High-speed confocal laser scanning microscopy using acousto-optic deflectors and a digital micromirror device

High-speed confocal laser scanning microscopy using acousto-...

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

作者： Bansal, V Patel, S Saggau, P Rice Univ Dept Bioengn Houston TX 77251 USA

ISBN: (纸本)0819452327

Neurons are known to possess active computational properties. To investigate these properties, it is desirable to study the electrical and chemical properties not only at a living neuron's cell body, but also at many sites within its dendritic arborization. However, currently available recording techniques force a tradeoff between spatial and temporal resolution. To overcome these limitations, we have developed a confocal microscope that can make multisite optical recordings at an effective frame rate that is sufficient to measure fast neuronal events, such as action potentials, that occur on a timescale of milliseconds. We accomplished this by combining acousto-optic deflectors for addressable point illumination with a digital micromirror device for addressable point detection. After developing a registration algorithm to ensure synchronicity between point illumination and point detection, we used light-scattering test preparations to demonstrate that our system is capable of optical sectioning and therefore capable of imaging in living brain tissue. Furthermore, we have shown that fluorescence changes can be monitored at an effective frame rate of 25 kHz.

关键词： confocal acousto-optic deflector AOD digital micromirror device DMD neuron

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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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Focus-tunable lens in limited-angle holographic tomography 24

Focus-tunable lens in limited-angle holographic tomography

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

作者： Kus, A. Krauze, W. Kujawinska, M. Warsaw Univ Technol Inst Micromech & Photon Sw Boboli 8 St PL-02525 Warsaw Poland

ISBN: (数字)9781510605824

ISBN: (纸本)9781510605817;9781510605824

In this paper a new, hardware-based solution for extending the depth of field in holographic tomography is presented. The solution is based on a 4f system and an electric, focus-tunable lens, which provides fast, motion-free defocusing of the plane conjugate with the camera, which acquires holograms. The optimum parameters for the required axial scanning are provided for a specific model of a commercially available tunable lens. Then, the quality of the system equipped with the designed module is analyzed and the reconstruction of a standard object (microsphere) scanned by the 4f-based defocusing system is presented. Finally, the result of the increased depth of field in the measurement domain is demonstrated with a reconstruction of a mouse fibroblast cell.

关键词： Tomographic phase microscopy digital holography diffraction tomography quantitative phase imaging 3D imaging refractive index

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