Principles of Fluorescence Spectroscopy

PRINCIPLES OF FLUORESCENCE SPECTROSCOPY 795
Sensing
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microscopy/single-molecule fluorescence spectroscopy.
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Single-Molecule Detection
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under ambient conditions: methodology and technology.
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Spectroscopy of Single Molecules
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Theory of Single-Molecule Fluorescence
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Time-Resolved Studies of Single Molecules
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PROBLEMS
P23.1. Suppose you have a fluorophore with a molar extinction
of 104,000 M –1 cm –1 at 600 nm, and a lifetime of
4 ns. Assume there is no intersystem crossing or blinking.
Calculate the light intensity in watts/cm 2 needed to
result in half of the molecule in the excited state. If the
illuminated is 1 µm 2 what is the needed power?
Assume the objective transmits 100% of the light.