Nature Methods 2, 910 - 919 (2005)
Published online: 18 November 2005; | doi:10.1038/nmeth817
Fluorescence microscopyJeff W Lichtman1
& José-Angel Conchello2, 31
Department of Molecular and Cell Biology, Harvard University, 7 Divinity Avenue, Cambridge, Massachusetts 02138, USA. 2
Molecular, Cell and Developmental Biology, Oklahoma Medical Research Foundation, 825 N.E. 13th Street, Oklahoma City, Oklahoma 73104, USA. 3
Program in Biomedical Engineering, University of Oklahoma, 100 East Boyd Street, Norman, Oklahoma 73019, USA.
Correspondence should be addressed to Jeff W Lichtman jeff@mcb.harvard.edu Although fluorescence microscopy permeates all of cell and molecular biology, most biologists have little experience with the underlying photophysical phenomena. Understanding the principles underlying fluorescence microscopy is useful when attempting to solve imaging problems. Additionally, fluorescence microscopy is in a state of rapid evolution, with new techniques, probes and equipment appearing almost daily. Familiarity with fluorescence is a prerequisite for taking advantage of many of these developments. This review attempts to provide a framework for understanding excitation of and emission by fluorophores, the way fluorescence microscopes work, and some of the ways fluorescence can be optimized.
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