Metabolic Profiling of Single Cancer Cells Using Mitochondrial ATP Probes

STAR Protoc. 2020 Jun 8;1(2):100048. doi: 10.1016/j.xpro.2020.100048. eCollection 2020 Sep 18.

Abstract

The metabolic activity of cells is interrelated with cell signaling, functions, and fate. Uncontrolled cancer cell proliferation requires metabolic adaptations. Research focusing on understanding the characteristics of cell metabolism is crucial for the development of novel diagnostic and therapeutic strategies. Here, we describe protocols for the ATP profiling of single cancer cells by fluorescence live-cell imaging. In response to distinct metabolic inhibitions, we record individual mitochondrial ATP dynamics using established Förster resonance energy transfer-based genetically encoded fluorescent ATP probes. For complete details on the use and execution of this protocol, please refer to Depaoli et al. (2018).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate* / analysis
  • Adenosine Triphosphate* / chemistry
  • Adenosine Triphosphate* / metabolism
  • Animals
  • Cell Line, Tumor
  • Fluorescence Resonance Energy Transfer
  • Fluorescent Dyes* / analysis
  • Fluorescent Dyes* / chemistry
  • Fluorescent Dyes* / metabolism
  • Humans
  • Mitochondria* / chemistry
  • Mitochondria* / metabolism
  • Molecular Imaging
  • Rats
  • Single-Cell Analysis / methods*
  • Tumor Cells, Cultured* / cytology
  • Tumor Cells, Cultured* / metabolism

Substances

  • Fluorescent Dyes
  • Adenosine Triphosphate