Peter J. Verveer's Advanced Fluorescence Microscopy: Methods and Protocols PDF

By Peter J. Verveer

ISBN-10: 1493920790

ISBN-13: 9781493920792

ISBN-10: 1493920804

ISBN-13: 9781493920808

This quantity presents an summary of complex fluorescence microscopy, masking a extensive variety of equipment. each one bankruptcy makes a speciality of a distinct strategy and gives a pragmatic advisor for software in organic platforms. Written within the hugely winning Methods in Molecular Biology sequence structure, chapters comprise introductions to their respective issues, lists of the mandatory fabrics and reagents, step by step, conveniently reproducible laboratory protocols, and pointers on troubleshooting and heading off identified pitfalls.

Authoritative and state of the art, Advanced Fluorescence Microscopy: tools and Protocols seeks to supply scientists with tools for organic structures which are of curiosity.

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Additional resources for Advanced Fluorescence Microscopy: Methods and Protocols

Example text

2 Two-Photon Excitation Microscopy Besides the discovery of fluorescent proteins, perhaps the most significant advance in bio-imaging of the last 25 years was the invention, in 1990, of two-photon excitation (2PE) microscopy [7]. Since then, 2PE has become widely popular for in vivo imaging of neuronal structure because of its superior depth penetration and reduced photobleaching compared to confocal or epifluorescence microscopy [8]. Imaging deeper with 2PE is achieved by means of excitation with an infrared laser and optics that collect much of the scattered emitted light, while maintaining excellent spatial resolution.

These novel geometries include the use of Bessel beams [90] and light sheets [91] to achieve thin 2PE planes perpendicular to the emission path or the implementation of spatial light modulators that shape the laser beam into an arbitrary light pattern, which allows for simultaneous imaging (or uncaging) at different locations [92] (see Note 4). Further developments in years to come will no doubt bring us closer to our goal of being able to record, with millisecond precision, the firing of thousands of neurons, distributed over a volume of brain tissue in behaving animals.

Graham-Smith SF, King TA (2000) Optics and photonics: an introduction. Millis BA (2012) Evanescent-wave field imaging: an introduction to total internal reflection fluorescence microscopy. Rappoport JZ, Simon SM (2009) Endocytic trafficking of activated EGFR is AP-2 dependent and occurs through preformed clathrin spots. Born M, Wolf E, Bhatia AB (1999) Principles of optics: electromagnetic theory of propagation, interference and diffraction of light. Sund SE, Swanson JA, Axelrod D (1999) Cell membrane orientation visualized by polarized total internal reflection fluorescence.

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Advanced Fluorescence Microscopy: Methods and Protocols by Peter J. Verveer


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