Characterization of a confocal microscope for time-resolved photon counting fluorescence |
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Authors: | Laetitia Schoutteten Patrick Denjean Robert B Pansu |
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Institution: | (1) ENS Cachan, PPSM (Photophysique et Photochimie Supramoléculaires et Macromoléculaires), URA 1906 CNRS, 61 avenue du Pré sident Wilson, 94235 Cachan Cedex, France |
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Abstract: | A confocal microscope setup is developed for time-resolved fluorescence measurements. It is added to a traditional cuvette
time-resolved setup, with a pumped Ti-Sa light source. The temporal resolution of 37 ps (FWHM) is not degraded, in comparison
with the cuvette setup also described. These setups allow both decay lifetime and anisotropy relaxation time determination.
Fluorescence correlation spectroscopy (FCS) is used to determine the observation point size. When associated with the calcium
probe calcium green, calcium concentration in single cells can be determined in 10 ms by simultaneous acquisition of early
and late fluorescence photons. |
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Keywords: | Time-resolved fluorescence confocal microscopy anisotropy fluorescence correlation spectroscopy calcium probes laser |
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