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Fluorophores for Confocal Microscopy

Photophysics and Photochemistry

  • Chapter
Handbook of Biological Confocal Microscopy

Abstract

Fluorescence is probably the most important optical readout mode in biological confocal microscopy, because it can be so much more sensitive and specific than absorbance or reflectance, and because it works so well with epi-illumination, which greatly simplifies scanner design. These advantages of fluorescence are critically dependent on the availability of suitable fluorophores that can either be tagged onto biological macromolecules to show their location, or whose optical properties are sensitive to the local environment. Despite the pivotal importance of good fluorophores, little is known about how to rationally design good ones. Whereas the concept of confocal microscopy is only a few decades old and nearly all the optical, electronic, and computer components to support it have been developed or redesigned in the last few years, the most popular fluorophores were developed more than a century ago (in the case of fluoresceins or rhodamines) or several billion years ago (in the case of phycobiliproteins). Moreover, whereas competition between commercial makers of confocal microscopes stimulates ardent efforts to refine the instrumentation, relatively few companies or academic scientists are interested in improving fluorophores.

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Authors and Affiliations

  1. Howard Hughes Medical Institute and Department of Pharmacology 0647, School of Medicine, University of California, San Diego, California, 92093, USA

    Roger Y. Tsien

  2. Department of Biological Sciences and Center for Light Microscope Imaging and Biotechnology, Carnegie Mellon University, Pittsburgh, Pennsylvania, 15213, USA

    Alan Waggoner

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  1. Roger Y. Tsien
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  1. Zoology Department, University of Wisconsin-Madison, Madison, Wisconsin, USA

    James B. Pawley

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Tsien, R.Y., Waggoner, A. (1995). Fluorophores for Confocal Microscopy. In: Pawley, J.B. (eds) Handbook of Biological Confocal Microscopy. Springer, Boston, MA. https://doi.org/10.1007/978-1-4757-5348-6_16

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