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Confocal fluorescence microscopy using spectral and lifetime information to simultaneously record four fluorophores with high channel separation
Author(s) -
CARLSSON K.,
LILJEBORG A.
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.1365-2818.1997.1450705.x
Subject(s) - confocal , fluorophore , microscope , fluorescence , confocal microscopy , detector , channel (broadcasting) , microscopy , optics , materials science , optical microscope , signal to noise ratio (imaging) , signal (programming language) , analytical chemistry (journal) , chemistry , physics , scanning electron microscope , computer science , chromatography , telecommunications , programming language
We demonstrate the possibility to increase substantially the number of simultaneously detected fluorophores by utilizing both spectral and lifetime information. Using a two‐detector confocal scanning laser microscope, experiments confirm that four different fluorophores can be detected with good channel separation. The signal‐to‐noise ratio (SNR) of the recorded images is investigated both theoretically and experimentally. It is found that in order to obtain a high SNR fluorophore lifetimes should differ by approximately an order of magnitude.