Low-cost, frequency-domain, fluorescence lifetime confocal microscopy

J. Microsc., Vol. 214, Pt. 1, April 2004, 36-42.

We describe the theory and implementation of a frequencydomain fluorescence lifetime confocal microscope using switcheddiode laser illumination. Standard, communications-type, radiofrequency electronics are used to provide inexpensive modulation references and to perform phase-sensitive detection. Thisallows the rapid acquisition of fluorescence intensity and lifetimeimages and their display in real time. We show fluorescencelifetime images of bead objects and fluorescence lifetime imagesof biological specimens from a single confocal scan.

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