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丁香实验推荐阅读
Confocal Microscopy on Xenopus laevis Oocytes and Embryos

There are many problems affecting microscopy in Xenopus. Xenopus oocytes and embryos are fragile and lack connective tissue. They are also full of yolk platelets, which prevents frozen sectioning because the yolk crystallizes and tears the sections. In addition, the yolk autofluoresc ...

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Confocal Imaging of Living Cells in Intact Embryos

How individual cells and tissues organize and coordinate to form the structure of a developing embryo is a major question in developmental biology. Equally intriguing is how cells in adult tissue respond during wound healing and tissue regeneration. In these morphogenetic processes th ...

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Live Confocal Analysis with Fluorescently Labeled Proteins

The development of laser scanning confocal microscopy provides a powerful means to observe structures and components within the cell. Equally important advances have also occurred in the development of reagents and techniques for generating functional fluorescently tagged p ...

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Fluorescent Calcium Indicators:Subcellular Behavior and Use in Confocal Imaging

Most animal cells maintain a large gradient of free calcium across the plasma membrane (10,000-fold or more), with intracellular free calcium held at a level of about 100 nM. To accomplish this, cells have an array of molecular machinery including ATP-driven calcium pumps, calcium exchangers, ...

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Live Imaging with Green Fluorescent Protein

If developmental biologists were given the chance to design the perfect cell-or tissue-specific marker, they would ensure that it had several properties. First, it would function in living animals, eliminating the need for fixation and dehydration and their associated artefacts. Sec ...

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Intracellular pH and pCa Measurement

Recent improvements in confocal technology permit the use of a confocal microscope as an effective tool to both measure concentration and visualize distribution of several ions. Effective discrimination against out-of-focus information in the confocal microscope permits for ...

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Measuring Dynamic Cell Volume In Situ by Confocal Microscopy

Regulation of cell volume is a fundamental homeostatic mechanism in the face of osmotic stress (1,2). One approach to understanding aspects of cell volume regulation involves the removal of cells from the matrix and manipulation in culture (3). Measurements are often also needed from cells w ...

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Imaging Thick Tissues with Confocal Microscopy

Confocal laser scanning microscopy (CLSM) can be used to obtain optical sections of thick tissues that are relatively free of interfering autofluorescence, and that do not strongly scatter or absorb either the excitation or emission light. This chapter provides protocols used to exami ...

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An Introduction to Confocal Imaging

The major application of confocal microscopy in the biomedical sciences is for imaging either fixed or living tissues that have usually been labeled with one or more fluorescent probes. When these samples are imaged using a conventional light microscope, the fluorescence in the specimen a ...

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Measurement in the Confocal Microscope

Accurate alignment of the microscope is vital for most measurements. Details of the adjustments will vary from system to system but the principles of alignment are universal. The methods given here should be appropriate for any spot-scanning confocal microscope but will not be directly ap ...

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Presentation of Confocal Images

Confocal microscopy is routinely used to produce high-resolution images of single, double-, and triple labeled fluorescent samples. The images are collected as single optical sections (2D imaging), as Z-series (3D imaging), as time-lapse series (2D over time), or as Z-series over time (3D over ...

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The Preparation of Stereoscopic 3D Illustrations of Confocal Data Sets for Publications and Slides

In this chapter we describe a simple method to prepare 3D illustrations from stereo-pairs of images from confocal sections. Methods for preparing both stereo slides and stereo images for publication using Microsoft’s PowerPoint� 97 and CorelDraw� 8 are described. The parallel and cross- ...

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Information Management of Confocal Microscopy Images: Traditional Text-Based Databases and Image Gallery Databases

Imaging methods, whether they use film, videotape, or digital capture methods, are only of use if you are able to readily categorize and retrieve the information and images that are produced. Modern imaging techniques are capable of generating large numbers of images of various dimensions and ...

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Morphing Confocal Images and Digital Movie Production

Visualizing change over time is essential to understanding biological events such as embryogenesis and development. More and more specimens can be visualized in the living state due to improvements in confocal microscopy, and the development of novel probes such as the green fluoresce ...

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Practical Considerations for Collecting Confocal Images

Conventional microscopy delivers two-dimensional images in real time and real color to the eye of the user. Confocal microscopy adds a third dimension by imaging only one plane within the sample at a time so that variations in depth can be quantified (1). This has both positive and negative aspects. T ...

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Imaging Gene Expression Using Antibody Probes

The merger of multiple immunoflurescent labeling and the laser scanning confocal microscope (LSCM) has greatly enhanced experimentation in many areas of biomedical research. Recently, genetic tools have become available for marking individual cells, or cells within a tissue of ge ...

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Fluorescent Probes for Confocal Microscopy

This chapter differs from others in this volume in that it does not describe protocols per se. Rather, it consists of checklists, cautions, tips, rules of thumb, and advice related to fluorescent probes in general and probes for confocal microscopy in particular.

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Single and Double FISH Protocols for Drosophila

In situ hybridization within whole-mount Drosophila tissues was made routine with the introduction of digoxigenin labeled probes and alkaline phosphatase based detection methods (1). However, this method of detection until recently has been limited by the required use of alkaline p ...

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Preparation of Yeast Cells for Confocal Microscopy

Confocal scanning microscopy has been successfully used for immunofluorescence work in yeast. The major axis of a Saccharomyces cerevisiae haploid cell is approx 4 μm. Optical sections of approximately 2� 0.4 μm thickness from fluorescently-labeled yeast cells can be obtained using t ...

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Confocal Microscopy of Plant Cells

The increasing availability of confocal microscopy has begun a revolution in plant biology in which microscopy has again become a powerful tool for understanding structure and function. Examples of applications include: three-dimensional (3D) reconstruction of the interphase ...

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