Format

Send to

Choose Destination
Cytometry A. 2014 Dec;85(12):1011-9. doi: 10.1002/cyto.a.22573. Epub 2014 Oct 1.

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

Author information

1
Baxter Laboratory for Stem Cell Biology, Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California; Divisions of Hematology and Oncology, Department of Medicine, Stanford University School of Medicine, Stanford, California.

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.

KEYWORDS:

fluorescent cellular barcoding; mass cytometry; mass-tag cellular barcoding; permeabilization; saponin

PMID:
25274027
PMCID:
PMC4361015
DOI:
10.1002/cyto.a.22573
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Wiley Icon for PubMed Central
Loading ...
Support Center