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Fluorescence discrimination in a reflecting environment
Author(s) -
Galassi L.
Publication year - 1990
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 - 10.1111/j.1365-2818.1990.tb02957.x
Subject(s) - fluorescence , optics , intensity (physics) , filter (signal processing) , beam (structure) , materials science , light beam , light intensity , physics , computer science , computer vision
SUMMARY A method to discriminate fluorescence from reflected or scattered light in epifluorescence microscopy is described. The method is based on the fact that the intensity of fluorescence measured with a long pass (LP) filter placed along the emerging beam (i.e. as a barrier filter) is higher than that measured with the same filter placed ahead of the objective (i.e. along the incident beam). On the other hand, when specific precautions are taken, the intensity of reflected or scattered light is identical to within a small difference. Within the margins of error of this small difference, it can be concluded that when the light intensity I measured with an LP filter along the emerging beam is higher than the intensity I ' measured with the same filter along the incident beam, a fluorescent emission is present and the fraction of fluorescent light in the emerging beam is at least ( I — I')/I.