Transient partial permeabilization with saponin enables cellular barcoding prior to surface marker staining

Year: 2014

Type: article

Abstract: Fluorescent cellular barcoding and mass-tag cellular barcoding are cytometric methods that enable high sample throughput, minimize inter-sample variation, and reduce reagent consumption. Previously employed barcoding protocols require that barcoding be performed after surface marker staining, complicating combining the technique with measurement of alcohol-sensitive surface epitopes. This report describes a method of barcoding fixed cells after a transient partial permeabilization with 0.02% saponin that results in efficient and consistent barcode staining with fluorescent or mass-tagged reagents while preserving surface marker staining. This approach simplifies barcoding protocols and allows direct comparison of surface marker staining of multiple samples without concern for variations in the antibody cocktail volume, antigen-antibody ratio, or machine sensitivity. Using this protocol, cellular barcoding can be used to reliably detect subtle differences in surface marker expression.

Source: Cytometry Part A

DOI: 10.1002/cyto.a.22573

Authors: Gregory K. Behbehani, Colin Thom, Eli R. Zunder, Rachel Finck, Brice Gaudilliere, Gabriela K. Fragiadakis, Wendy J. Fantl, Garry P. Nolan

Institutions: Stanford Medicine, Stanford University

Language: English

Topic: Single-cell and spatial transcriptomics

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: bronze

Funders: Bill and Melinda Gates Foundation, California Institute for Regenerative Medicine, European Commission, National Institutes of Health, U.S. Public Health Service, National Heart, Lung, and Blood Institute, National Cancer Institute, National Institute of Allergy and Infectious Diseases

OpenAlex ID: W1537768706