The fluorescent toolbox for assessing protein location and function.
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.
Giepmans
Ben N G
BN
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.
Adams
Stephen R
SR
Ellisman
Mark H
MH
Tsien
Roger Y
RY
D004058:Diffusion; D004798:Enzymes / Q000378:metabolism; D005453:Fluorescence; D031541:Fluorescence Resonance Energy Transfer; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes / Q000737:chemistry; D005821:Genetic Techniques; D007150:Immunohistochemistry; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics; D008854:Microscopy, Electron; D015336:Molecular Probe Techniques; D011487:Protein Conformation; D021381:Protein Transport; D011506:Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D045663:Quantum Dots
D004798:Enzymes; D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D011506:Proteins