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Structure‐orientated fluorescence photobleaching analysis: a method for double fluorescent labelling studies [Note 1. Presented at 3D Imaging Sciences in Microscopy, Oxford, 14–17 ...]
Author(s) -
GHAUHARALI R. I.,
VAN DRIEL R.,
BRAKENHOFF G. J.
Publication year - 1997
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1046/j.1365-2818.1997.d01-630.x
Subject(s) - photobleaching , photodegradation , fluorophore , fluorescence , quenching (fluorescence) , chemistry , labelling , photochemistry , fluorescence recovery after photobleaching , biological system , biophysics , materials science , optics , physics , biochemistry , biology , catalysis , photocatalysis
Differences in the degree of photodegradation can be used for fluorophore identification in double fluorescently labelled specimens. Based on the use of morphological information, a noise‐insensitive method is presented for discriminating between the fluorophores, assuming spatially uniform photodegradation. Separate images of the labelled structures can be obtained. Alternatively, with spatially nonuniform photodegradation, the photodynamics of one fluorophore — i.e. photodegradation, concentration associated quenching, etc. — in relation to its microenvironment can be investigated.