Characterizing the Electron Transport Chain: Functional Approach Using Extracellular Flux Analyzer on Mouse Tissue Samples

Ting Liang, Jay Dunn, Xin Zou, Bijaya Nayak, Yuji Ikeno, Lihong Fan, Yidong Bai

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

The Seahorse Extracellular Flux Analyzer enables the high-throughput characterization of oxidative phosphorylation capacity based on the electron transport chain organization and regulation with relatively small amount of material. This development over the traditional polarographic Clark-type electrode approaches make it possible to analyze the respiratory features of mitochondria isolated from tissue samples of particular animal models. Here we provide a description of an optimized approach to carry out multi-well measurement of O2 consumption, with the Agilent Seahorse XFe96 analyzer on mouse brain and muscles to determine the tissue-specific oxidative phosphorylation properties. Protocols include the preparation of the tissue samples, isolation of mitochondria, and analysis of their function; in particular, the preparation and optimization of the reagents and samples.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press
Pages117-128
Number of pages12
DOIs
StatePublished - 2022

Publication series

NameMethods in Molecular Biology
Volume2497
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Brain
  • Electron transport chain
  • Muscle
  • O consumption
  • Seahorse

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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