Ludwig-Maximilians-Universität, Chair of Metabolic Biochemistry

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Two postprocessing techniques for the elimination of background autofluorescence for fluorescence lifetime imaging microscopy

J Biomed Opt. 2008 Jan-Feb;13(1):014008.

Authors/Editors: Jones PB
Rozkalne A
Meyer-Luehmann M
Spires-Jones TL
Makarova A
Kumar AT
Berezovska O
Bacskai BB
Hyman BT
Publication Date: 2008
Type of Publication: Journal Article
The analysis of fluorescence lifetime imaging microscopy (FLIM) data under complex biological conditions can be challenging. Particularly, the presence of short-lived autofluorescent aggregates can confound lifetime measurements in fluorescence energy transfer (FRET) experiments, where it can become confused with the signal from exogenous fluorophores. Here we report two techniques that can be used to discriminate the contribution of autofluorescence from exogenous fluorphores in FLIM. We apply the techniques to transgenic mice that natively express yellow fluorescence protein (YFP) in a subset of cortical neurons and to histological slices of aged human brain tissue, where we study the misfolding of intracellular tau protein in the form of neurofibrillary tangles.

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