Developing Analysis Protocols for Monitoring Intracellular Oxygenation Using Fluorescence Lifetime Imaging of Myoglobin-mCherry.

ABSTRACT: Oxygen (O₂) is a critical metabolite for cellular function as it fuels aerobic cellular metabolism; further, it is a known regulator of gene expression. Monitoring oxygenation within cells and organelles can provide valuable insights into how O₂, or lack thereof, both influences and responds to cell processes. In recent years, fluorescence lifetime imaging microscopy (FLIM) has been used to track several probe concentration independent intracellular phenomena, such as pH, viscosity, and, in conjunction with Förster resonance energy transfer (FRET), protein-protein interactions. Here, we describe methods for synthesizing and expressing the novel FLIM-FRET intracellular O₂ probe Myoglobin-mCherry (Myo-mCherry) in cultured cell lines, as well as acquiring FLIM

SUBMITTER: Alspaugh G 

PROVIDER: S-EPMC8813554 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature