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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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Accurate cell counts in live mouse embryos using optical quadrature and differential interference contrast microscopy

Accurate cell counts in live mouse embryos using optical qua...

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

作者： Warger, William C., II Newmark, Judith A. Zhao, Bing Warner, Carol M. DiMarzio, Charles A. Northeastern Univ Dept Elect & Comp Engn 440 Dana Bldg360 Huntington Ave Boston MA 02115 USA Northeastern Univ Dept Biol Boston MA 02115 USA

ISBN: (纸本)0819461326

Present imaging techniques used in in vitro fertilization (IVF) clinics are unable to produce accurate cell counts in developing embryos past the eight-cell stage. We have developed a method that has produced accurate cell counts in live mouse embryos ranging from 13 - 25 cells by combining Differential Interference Contrast (DIC) and Optical Quadrature microscopy. Optical Quadrature microscopy is an interferometric imaging modality that measures the amplitude and phase of the signal beam that travels through the embryo. The phase is transformed into an image of optical path length difference, which is used to determine the maximum optical path length deviation of a single cell. DIC microscopy gives distinct cell boundaries for cells within the focal plane when other cells do not lie in the path to the objective. Fitting an ellipse to the boundary of a single cell in the DIC image and combining it with the maximum optical path length deviation of a single cell creates an ellipsoidal model cell of optical path length deviation. Subtracting the model cell from the Optical Quadrature image will either show the optical path length deviation of the culture medium or reveal another cell underneath. Once all the boundaries are used in the DIC image, the subtracted Optical Quadrature image is analyzed to determine the cell boundaries of the remaining cells. The final cell count is produced when no more cells can be subtracted. We have produced exact cell counts on 5 samples, which have been validated by Epi-Fluorescence images of Hoechst stained nuclei.

关键词： Optical Quadrature microscopy Differential Interference Contrast embryo viability phase subtraction cell counting method IVF OQM QTM

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Closing the spectral gap - the transition from fixed-parameter fluorescence to tunable devices in confocal microscopy.

Closing the spectral gap - the transition from fixed-paramet...

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

作者： Borlinghaus, Rolf Gugel, Hilmar Albertano, Patrizia Seyfried, Volker Leica Microsyst CMS Mannheim Germany Univ Rome Dept Biol Rome Italy

ISBN: (纸本)0819461326

Modem microscopy in life sciences is ruled by development and exploration of new dyes and stains (probes for histochemical staining, quantum dots, fluorescent proteins etc.) on one side, and technological improvements and innovations for fluorescence microscopy - especially high resolution and optical sectioning microscopy - on the other side. Concerning the technical innovations, several ingenious inventions have been made available for confocal microscopy. First, the acousto optical tunable filter, which allows switching and dimming of laser lines. Second the spectral detector, employing mirror sliders in front of the detectors which allow continuous tuning of the spectral emission band detected by the sensor. Third, the most challenging task: a substitute to the classical beam splitter - the device which is restricting fluorescence microscopy most. This was solved by introduction of the acousto optical beam splitter. The very last device which is still lacking flexibility is the laser source, operating only at non-equidistant frequencies and requiring a set of quite different laser sources as gas lasers, solid state lasers or diode lasers. A new approach by supercontinuum light sources is presented and discussed, which significantly enhances flexibility and coverage of the excitation spectra of typical, rare and natural fluorochromes.

关键词： confocal spectral white laser supercontinuum AOBS cyanobacteria excitation-emission correlation microscopy sensitivity multi-parameter fluorescence

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A narrowband dual-axes confocal reflectance microscope for distinguishing colonic neoplasia

A narrowband dual-axes confocal reflectance microscope for d...

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

作者： Liu, Jonathan T. C. Mandella, Michael J. Contag, Christopher H. Kino, Gordon S. Wang, Thomas D. Stanford Univ Ginzton Lab Box N-123450 Via Palou Stanford CA 94305 USA Stanford Univ Sch Med Stanford CA 94305 USA

ISBN: (纸本)0819461326

A dual-axes confocal reflectance microscope has been developed that utilizes a narrowband source at 1310 nm to achieve high axial resolution, image contrast, field of view, and tissue penetration for distinguishing among normal, hyperplastic, and dysplastic colonic mucosa ex vivo. Light is collected off-axis using a low numerical aperture objective to obtain vertical image sections, with 4 to 5-mu m resolution, at tissue depths up to 610 mu m. Post-objective scanning enables a large field of view (610 x 640 mu m) and balanced-heterodyne detection provides sensitivity to collect vertical sections at two frames per second. System optics are optimized to effectively reject out-of-focus scattered light without use of a low-coherence gate. This design is scalable to millimeter dimensions, and the results demonstrate the potential for a miniature instrument to detect pre-cancerous tissues, and hence to perform in vivo histopathology.

关键词： laser scanning confocal microscopy in vivo imaging reflectance microscopy heterodyne detection 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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New techniques in confocal microscopy

New techniques in confocal microscopy

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Laser Science, LS 2006

作者： Mertz, Jerome Boston University Biomedical Engineering Department 44 Cummington St. Boston MA 02215 United States

Two new optical techniques are presented that lead to out-of-focus background rejection. The first is autoconfocal microscopy, which reveals phase-gradients. The second is dynamic speckle illumination microscopy, whic... 详细信息

ISBN: (纸本)1557528187

关键词： image processing Imaging techniques Laser speckle Speckle patterns Structured illumination microscopy Three dimensional imaging Imaging systems (110.0110) Three-dimensional image acquisition (110.6880) microscopy (180.0180) Confocal microscopy (180.1790)

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Space Imaging Measurement System based on fixed lens and moving detector

Space Imaging Measurement System based on fixed lens and mov...

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conference on Novel Optical Systems Design and Optimization ix

作者： Akiyama, Akira Doshida, Minoru Mutoh, Eiichiro Kumagai, Hideo Yamada, Hirofumi Ishii, Hiromitsu Kanazawa Tech Coll 2-270 Hisayasu Kanazawa Ishikawa 9218601 Japan Japan Def Agcy Tech Res & Dev Inst Tachikawa Tokyo 1908533 Japan Kawasaki Heavy Ind Co Ltd Aerosp Co Minato Ku Tokyo 1056116 Japan Tamagawa Seiki Co Guidance & Nav Sect Nagano 3958515 Japan Kanazawa Tech Coll Kanazawa Ishikawa 9218601 Japan Nihon Univ Dept Elect Engn Coll Sci & Technol Chiyoda Ku Tokyo 1018308 Japan

ISBN: (纸本)081946368X

We have developed the Space Imaging Measurement System based on the fixed lens and fast moving detector to the control of the autonomous ground vehicle. The space measurement is the most important task in the development of the autonomous ground vehicle. In this study we move the detector back and forth along the optical axis at the fast rate to measure the three-dimensional image data. This system is just appropriate to the autonomous ground vehicle because this system does not send out any optical energy to measure the distance and keep the safety. And we use the digital camera of the visible ray range. Therefore it gives us the cost reduction of the three-dimensional image data acquisition with respect to the imaging laser system. We can combine many pieces of the narrow space imaging measurement data to construct the wide range three-dimensional data. This gives us the improvement of the image recognition with respect to the object space. To develop the fast movement of the detector, we build the counter mass balance in the mechanical crank system of the Space Imaging Measurement System. And then we set up the duct to prevent the optical noise due to the ray not coming through lens. The object distance is derived from the focus distance which related to the best focused image data is selected from the image of the maximum standard deviation in the standard deviations of series images.

关键词： Sensors Imaging systems Modulation transfer functions CCD cameras Geometrical optics 3D image processing CCD image sensors Distance measurement image sensors Ranging

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Establishing multi-modality datasets with the incorporation of 3D histopathology for soft tissue classification

Establishing multi-modality datasets with the incorporation ...

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Medical Imaging 2006 conference

作者： de Ryk, Jessica Thiesse, Jacqueline Reinhardt, Joseph M. Hoffman, Eric A. McLennan, Geoffrey Univ Iowa 200 Hawkins Dr Iowa City IA 52242 USA

ISBN: (纸本)0819464236

The development of multi-modality image analysis has gained increasing popularity over recent years. Multi-modality image databases are being developed to benefit patient clinical care, research and education. The incorporation of histopathology in these multi-modality datasets is complicated by the large differences in image quality, content and spatial association. We have developed a novel system, the large-scale image microtome array (LIMA), to bridge the gap between non-structurally destructive and destructive imaging such that reliable registration and incorporation of three-dimensional (3D) histopathology can be achieved. We have developed registration algorithms to align the micro-CT, LIMA and histopathology data to a common coordinate system. Using this multi-modality image dataset we have developed a classification algorithm to identify on a pixel basis, the tissue types present. The output from the classification processing is a 3D color coded map of tissue distributions. The resulting complete dataset provides an abundance of valuable information relating to the tissue sample including density, anatomical structure, color, texture and cellular information in three dimensions. In this study we have chosen to use normal and diseased lung tissue, however the flexibility of the image acquisition and subsequent processing algorithms makes it applicable to any soft organ tissue.

关键词： 3D histopathology microscopy computed micro-tomography thin-plate spline registration classification

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Multi-dimensional microscopy and its application in investigating instant effects of radiations on living cells

Multi-dimensional microscopy and its application in investig...

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The 3rd Asian and Oceanian conference on Photobiology

作者： Yao-Xiong Huang Institute of Biomedical Engineering Ji Nan University Guang Zhou China.

＜正＞The present paper introduces our newly developed techniques of multi-dimensional microscopy for measuring the structure and functions of living cells, simultaneously at cellular, sub-cellular, and intracellular molecule levels. The technical system is based on an inverted fluorescent microscope, its information acquisition and processing is organized in 8-dimensions: 3 dimensional image is acquired by a 3 dimensional relief image construction system;the 4th dimension （wave length） is taken by a fast micro-spectrophotometry;which, accompanied with a dynamic image analyzing system, also gives the information in the 5th dimension （time） by giving the absorption spectra and images in a time-lapse mode;the 6th -8th dimensions are organized by the lights from the sample: the transmitted light, gives the information of image and absorption spectroscopy;the scattered light, gives the information about the dynamic parameters of intracellular molecule and the cell membrane, as well as the chemical structure and concentration of the molecules;the fluorescence, gives the information of the labeled molecules in the cell. Therefore, the multi-dimensional microscopy is capable of performing measurement on the parameters of single intact cells include: 1) the chemical structure and concentration as well as the hydrodynamic radius of the intracellular molecules;2) the 3-D morphology and molecular distribution of the cell;3) the functional change of the intracellular protein;4) the flexibility and rigidity of the cell membrane;5) the differentiation, proliferation and the activity of the cell.

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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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