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Simultaneous confocal lifetime imaging of multiple fluorophores using the intensity‐modulated multiple‐wavelength scanning (IMS) technique
Author(s) -
- Carlsson,
Liljeborg
Publication year - 1998
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.1365-2818.1998.00362.x
Subject(s) - confocal , optics , modulation (music) , confocal microscopy , intensity (physics) , wavelength , fluorescence , microscope , signal to noise ratio (imaging) , signal (programming language) , laser , materials science , intensity modulation , physics , phase modulation , computer science , phase noise , acoustics , programming language
We demonstrate the simultaneous recording of confocal lifetime images of multiple fluorophores. The confocal microscope used in the study combines intensity‐modulated laser illumination, lock‐in detection and spectral separation of the fluorescent light. A theoretical investigation is presented that describes how the signal‐to‐noise ratio (SNR) depends on various factors such as modulation frequency, degree of modulation and number of detected photons. Theory predicts that, compared with ordinary intensity images, lifetime images will have a SNR that is, at best, approximately four times lower. Experimental results are presented that confirm this prediction.