| Evanescent field excitation of fluorescence by epi-illumination microscopy. | |
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MedLine Citation:
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PMID: 20556034 Owner: NLM Status: In-Data-Review |
Abstract/OtherAbstract:
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By simple modification of the pattern of fluorescence excitation light in an epi-illumination inverted microscope, one can achieve conditions that produce total internal reflection fluorescence (TIRF) by evanescent wave excitation. Though traditionally requiring a collimated beam traversing through a special prism, TIRF also can be achieved by epi-illumination through the periphery of a 1.4 numerical aperture objective. An opaque disk of appropriate size is placed in the illumination path external to the microscope so as to cast a sharp, real-image shadow at the objective's back focal plane. This shadow allows a hollow cone of epiillumination rays traveling at only super-critical angles to reach the glass/water interface at the sample plane. Three kinds of TIRF illumination patterns can be produced by variations of this scheme: (1) a small spot of illumination of 1.5 microm radius by use of a laser light source, (2) a large region of illumination by use of a laser-illuminated diffusing screen located upbeam from the opaque disk, and (3) a large region of illumination by use of a conventional mercury arc. |
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Authors:
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A L Stout; D Axelrod |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Applied optics Volume: 28 ISSN: 0003-6935 ISO Abbreviation: Appl Opt Publication Date: 1989 Dec |
Date Detail:
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Created Date: 2010-06-17 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0247660 Medline TA: Appl Opt Country: United States |
Other Details:
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Languages: eng Pagination: 5237-42 Citation Subset: - |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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