Open Access
1 January 2008 Two postprocessing techniques for the elimination of background autofluorescence for fluorescence lifetime imaging microscopy
Phill B. Jones, Aneta Rozkalne, Melanie Meyer-Luehmann, Tara L. Spires-Jones, Alexandra Makarova, Anand T. N. Kumar, Oksana Berezovska, Brian J. Bacskai, Bradley T. Hyman
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Abstract
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.
©(2008) Society of Photo-Optical Instrumentation Engineers (SPIE)
Phill B. Jones, Aneta Rozkalne, Melanie Meyer-Luehmann, Tara L. Spires-Jones, Alexandra Makarova, Anand T. N. Kumar, Oksana Berezovska, Brian J. Bacskai, and Bradley T. Hyman "Two postprocessing techniques for the elimination of background autofluorescence for fluorescence lifetime imaging microscopy," Journal of Biomedical Optics 13(1), 014008 (1 January 2008). https://doi.org/10.1117/1.2837169
Published: 1 January 2008
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CITATIONS
Cited by 15 scholarly publications.
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KEYWORDS
Atrial fibrillation

Fluorescence lifetime imaging

Fluorescence resonance energy transfer

Proteins

Image segmentation

Microscopy

Luminescence

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