Best Practices for Preparing a Single Cell Suspension from Solid Tissues for Flow Cytometry.
disaggregation
flow cytometry
single cell suspension
tissue digestion
Journal
Cytometry. Part A : the journal of the International Society for Analytical Cytology
ISSN: 1552-4930
Titre abrégé: Cytometry A
Pays: United States
ID NLM: 101235694
Informations de publication
Date de publication:
02 2019
02 2019
Historique:
received:
03
08
2018
revised:
26
09
2018
accepted:
02
11
2018
pubmed:
14
12
2018
medline:
9
4
2020
entrez:
8
12
2018
Statut:
ppublish
Résumé
Preparing a single cell suspension is a critical step in any solid tissue flow cytometry experiment. Tissue dissection, enzymatic digestion, and mechanical dissociation are three significant steps leading to the degradation of the extracellular matrix and the isolation of single cells, allowing the generation of high-quality flow cytometry data. Cells and the extracellular matrix contain various proteins and other structures which must be considered when designing a tissue digestion protocol to preserve the viability of cells and the presence of relevant antigens while digesting matrix components and cleaving cell-cell junctions. Evaluation of the single cell suspension is essential before proceeding with the labeling of the cells as high viability and absence of cell debris and aggregates are critical for flow cytometry. The information presented should be used as a general guide of steps to consider when preparing a single cell suspension from solid tissues for flow cytometry experiments. © 2018 International Society for Advancement of Cytometry.
Identifiants
pubmed: 30523671
doi: 10.1002/cyto.a.23690
pmc: PMC6375754
mid: NIHMS997084
doi:
Substances chimiques
Antigens
0
Proteins
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
219-226Subventions
Organisme : NHLBI NIH HHS
ID : P01 HL081064
Pays : United States
Organisme : NHLBI NIH HHS
ID : P01 HL103453
Pays : United States
Organisme : NHLBI NIH HHS
ID : U10 HL109250
Pays : United States
Informations de copyright
© 2018 International Society for Advancement of Cytometry.
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