Using redox-sensitive fluorescent probes to record real-time reactive oxygen species production in cells from mouse carotid body slices

STAR Protoc. 2021 May 11;2(2):100535. doi: 10.1016/j.xpro.2021.100535. eCollection 2021 Jun 18.

Abstract

Reactive oxygen species (ROS) are important signaling molecules for physiologic processes such as acute response to hypoxia. However, reliable real-time ROS measurement in cells has been a long-standing methodological challenge. Here, we present a protocol to record acute changes in ROS production in sensory cells from mouse carotid body (CB) slices using redox-sensitive green fluorescent protein probes and microfluorimetry. This protocol provides sensitive and reproducible quantification of ROS during acute hypoxia in different subcellular compartments of CB glomus cells. For complete details on the use and execution of this protocol, please refer to Fernández-Agüera et al. (2015) and Arias-Mayenco et al. (2018).

Keywords: Cell Biology; Metabolism; Microscopy; Molecular/Chemical Probes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Carotid Body* / chemistry
  • Carotid Body* / metabolism
  • Female
  • Fluorescent Dyes* / analysis
  • Fluorescent Dyes* / metabolism
  • Fluorometry
  • Histocytochemistry
  • Male
  • Mice
  • Oxidation-Reduction
  • Reactive Oxygen Species* / analysis
  • Reactive Oxygen Species* / metabolism

Substances

  • Fluorescent Dyes
  • Reactive Oxygen Species