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

Leelatian N, Doxie DB, Greenplate AR, Sinnaeve J, Ihrie RA, Irish JM
Curr Protoc Mol Biol. 2017 118: 25C.1.1-25C.1.23

PMID: 28369679 · PMCID: PMC5518778 · DOI:10.1002/cpmb.37

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 >10 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. © 2017 by John Wiley & Sons, Inc.

Copyright © 2017 John Wiley & Sons, Inc.

MeSH Terms (6)

Cell Separation Cryopreservation Flow Cytometry Humans Neoplasms Single-Cell Analysis

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