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Single‐Molecule Microscopy of the Green Fluorescent Protein Using Simultaneous Two‐Color Excitation
Author(s) -
Jung Gregor,
Wiehler Jens,
Steipe Boris,
Bräuchle Christoph,
Zumbusch Andreas
Publication year - 2001
Publication title -
chemphyschem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.016
H-Index - 140
eISSN - 1439-7641
pISSN - 1439-4235
DOI - 10.1002/1439-7641(20010618)2:6<392::aid-cphc392>3.0.co;2-7
Subject(s) - green fluorescent protein , microscopy , fluorescence , brightness , fluorescence microscope , biophysics , chemistry , molecule , photobleaching , materials science , optics , biology , biochemistry , physics , organic chemistry , gene
In vivo microscopy of the Green Fluorescent Protein (GFP), the most important label in cell biology, with single‐molecule sensitivity is hampered by an insufficient signal‐to‐noise ratio. A significant improvement is obtained with a novel two‐color excitation technique. The picture clearly shows the increased brightness of GFP in in vitro single‐molecule assays and in live‐cell microscopy under two‐color illumination (upper cell) as compared to normal illumination (lower cell).