Time-Resolved Imaging of Mitochondrial Flavin Fluorescence and Its Applications for Evaluating the Oxidative State in Living Cardiac Cells

Methods Mol Biol. 2021:2275:403-414. doi: 10.1007/978-1-0716-1262-0_26.

Abstract

Time-resolved fluorescence spectrometry is a highly valuable technological tool to detect and characterize mitochondrial metabolic oxidative changes by means of endogenous fluorescence. Here, we describe detection and measurement of endogenous mitochondrial flavin fluorescence directly in living cardiac cells using fluorescence lifetime imaging microscopy (FLIM) after excitation with 473 nm picoseconds (ps) laser. Time-correlated single photon counting (TCSPC) method is employed.

Keywords: Endogenous flavin fluorescence; Energy metabolism; FLIM; Mitochondrial oxidative state; TCSPC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Energy Metabolism
  • Flavins / chemistry*
  • Microscopy, Fluorescence
  • Mitochondria, Heart / metabolism*
  • Myocytes, Cardiac / cytology*
  • Myocytes, Cardiac / metabolism
  • Oxidative Phosphorylation
  • Oxidative Stress
  • Rats
  • Spectrometry, Fluorescence

Substances

  • Flavins