A redox-based characterization of human immune cell subsets by polychromatic flow cytometry.
View/ Open
Volume
4
Pagination
102632 - ?
DOI
10.1016/j.xpro.2023.102632
Journal
STAR Protoc
Issue
Metadata
Show full item recordAbstract
Cellular redox state determinants are traditionally studied using fluorescent microscopy and immunoblot analysis; however, no procedure has been developed for simultaneous measurement in various immune cell subsets. Here, we present a flow cytometry assay for measuring antioxidant defense systems and reactive oxygen species simultaneously in T, B, and natural killer lymphocytes. We describe steps for preparing and treating peripheral blood mononuclear cells, surface and dye staining, cell fixation/permeabilization, and intracellular staining. We then detail machine standardization, acquisition, and analysis.