Fluorescence microscopy

Nature Methods volume 2, pages910–919(2005)Cite this article

Abstract

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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Figure 1: Fluorescence fundamentals.
Figure 2: The spectral and structural properties of fluorophores.
Figure 3: The fluorescence microscope.

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Author information

Affiliations

  1. Department of Molecular and Cell Biology, Harvard University, 7 Divinity Avenue, Cambridge, 02138, Massachusetts, USA

    Jeff W Lichtman

  2. Molecular, Cell and Developmental Biology, Oklahoma Medical Research Foundation, 825 N.E., 13th Street, Oklahoma City, 73104, Oklahoma, USA

    José-Angel Conchello

  3. Program in Biomedical Engineering, University of Oklahoma, 100 East Boyd Street, Norman, 73019, Oklahoma, USA

    José-Angel Conchello

Authors
  1. Jeff W Lichtman
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  2. José-Angel Conchello
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Corresponding author

Correspondence to Jeff W Lichtman.

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The authors declare no competing financial interests.

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Lichtman, J., Conchello, JA. Fluorescence microscopy. Nat Methods 2, 910–919 (2005). https://doi.org/10.1038/nmeth817

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