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Curr Protoc Mol Biol. 2017 Apr 3;118:25C.1.1-25C.1.23. doi: 10.1002/cpmb.37.

Preparing Viable Single Cells from Human Tissue and Tumors for Cytomic Analysis.

Author information

1
Department of Cancer Biology, Vanderbilt University, Nashville, Tennessee.
2
Department of Pathology, Microbiology and Immunology, Vanderbilt University, Nashville, Tennessee.
3
Department of Neurological Surgery, Vanderbilt University Medical Center, Nashville, Tennessee.
4
Department of Cell and Developmental Biology, Vanderbilt University, Nashville, Tennessee.

Abstract

Mass cytometry is a single-cell biology technique that samples >500 cells per second, measures >35 features per cell, and is sensitive across a dynamic range of >104 relative intensity units per feature. This combination of technical assets has powered a series of recent cytomic studies where investigators used mass cytometry to measure protein and phospho-protein expression in millions of cells, characterize rare cell types in healthy and diseased tissues, and reveal novel, unexpected cells. However, these advances largely occurred in studies of blood, lymphoid tissues, and bone marrow, since the cells in these tissues are readily obtained in single-cell suspensions. This unit establishes a primer for single-cell analysis of solid tumors and tissues, and has been tested with mass cytometry. The cells obtained from these protocols can be fixed for study, cryopreserved for long-term storage, or perturbed ex vivo to dissect responses to stimuli and inhibitors.

KEYWORDS:

cytometry; dissociation; human tissue; human tumor; single cell

PMID:
28369679
PMCID:
PMC5518778
DOI:
10.1002/cpmb.37
[Indexed for MEDLINE]
Free PMC Article

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