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Proceedings Paper

Optimization of analysis protocol and probe expression for monitoring intracellular oxygenation using fluorescence lifetime imaging of myoglobin-mCherry (Conference Presentation)
Author(s): Rozhin Penjweini; Greg Alspaugh; Branden Roarke; Alexandra Chand; Jay R. Knutson

Paper Abstract

Fluorescence lifetime imaging (FLIM) of Myoglobin (Myo)-mCherry, is used for sensing oxygen partial pressure (pO2) in the intracellular environment. Herein, we present the potential sources of lifetime error such as sample oversaturation or dimeric Myo-mCherry configurations resulting in self-quenching fluorophores. We also provide a correction protocol for Myo-mCherry expression, adjusting parameters to account for second harmonic generation (SHG) components and dark counts that result in accurate mean lifetime values and pO2 in the cellular environment.

Paper Details

Date Published: 9 March 2020
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Proc. SPIE 11244, Multiphoton Microscopy in the Biomedical Sciences XX, 112441T (9 March 2020); doi: 10.1117/12.2550660
Show Author Affiliations
Rozhin Penjweini, National Institutes of Health Clinical Ctr. (United States)
Greg Alspaugh, National Institutes of Health Clinical Ctr. (United States)
Branden Roarke, National Institutes of Health Clinical Ctr. (United States)
Alexandra Chand, National Institutes of Health Clinical Ctr. (United States)
Jay R. Knutson, National Institutes of Health Clinical Ctr. (United States)


Published in SPIE Proceedings Vol. 11244:
Multiphoton Microscopy in the Biomedical Sciences XX
Ammasi Periasamy; Peter T. C. So; Karsten König, Editor(s)

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