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Transient partial permeabilization with saponin enables cellular barcoding prior to surface marker staining
Author(s) -
Behbehani Gregory K.,
Thom Colin,
Zunder Eli R.,
Finck Rachel,
Gaudilliere Brice,
Fragiadakis Gabriela K.,
Fantl Wendy J.,
Nolan Garry P.
Publication year - 2014
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.22573
Subject(s) - staining , biology , dna barcoding , mass cytometry , barcode , computational biology , microbiology and biotechnology , gene , biochemistry , computer science , genetics , phenotype , ecology , operating system
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. © 2014 International Society for Advancement of Cytometry