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Science 312 (5771): 217-224

Copyright © 2006 by the American Association for the Advancement of Science

The Fluorescent Toolbox for Assessing Protein Location and Function

Ben N. G. Giepmans,1,2 Stephen R. Adams,2 Mark H. Ellisman,1 Roger Y. Tsien2,3*

Abstract: Advances in molecular biology, organic chemistry, and materials science have recently created several new classes of fluorescent probes for imaging in cell biology. Here we review the characteristic benefits and limitations of fluorescent probes to study proteins. The focus is on protein detection in live versus fixed cells: determination of protein expression, localization, activity state, and the possibility for combination of fluorescent light microscopy with electron microscopy. Small organic fluorescent dyes, nanocrystals ("quantum dots"), autofluorescent proteins, small genetic encoded tags that can be complexed with fluorochromes, and combinations of these probes are highlighted.

1 National Center for Microscopy and Imaging Research, Center for Research in Biological Systems, Department of Neurosciences, University of California, San Diego, La Jolla, CA 92093, USA.
2 Department of Pharmacology, University of California, San Diego, La Jolla, CA 92093, USA.
3 Department of Chemistry and Biochemistry and Howard Hughes Medical Institute, University of California, San Diego, La Jolla, CA 92093, USA.

* To whom correspondence should be addressed. E-mail: rtsien{at}

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