Optimization of analysis protocol and probe expression for monitoring intracellular oxygenation using fluorescence lifetime imaging of myoglobin-mCherry (Conference Presentation)

Rozhin Penjweini; Greg Alspaugh; Branden Roarke; Alexandra Chand; Jay R. Knutson

doi:10.1117/12.2550660

9 March 2020 Optimization of analysis protocol and probe expression for monitoring intracellular oxygenation using fluorescence lifetime imaging of myoglobin-mCherry (Conference Presentation)

Rozhin Penjweini, Greg Alspaugh, Branden Roarke, Alexandra Chand, Jay R. Knutson

Author Affiliations +

Proceedings Volume 11244, Multiphoton Microscopy in the Biomedical Sciences XX; 112441T (2020) https://doi.org/10.1117/12.2550660
Event: SPIE BiOS, 2020, San Francisco, California, United States

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.

Conference Presentation

Citation Download Citation

Rozhin Penjweini, Greg Alspaugh, Branden Roarke, Alexandra Chand, and Jay R. Knutson "Optimization of analysis protocol and probe expression for monitoring intracellular oxygenation using fluorescence lifetime imaging of myoglobin-mCherry (Conference Presentation)", Proc. SPIE 11244, Multiphoton Microscopy in the Biomedical Sciences XX, 112441T (9 March 2020); https://doi.org/10.1117/12.2550660

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