The modified FACS calcein AM retention assay: A high throughput flow cytometer based method to measure cytotoxicity.
J Immunol Methods
; 434: 16-23, 2016 07.
Article
em En
| MEDLINE
| ID: mdl-27084117
ABSTRACT
Current methods to determine cellular cytotoxicity in vitro are hampered by background signals that are caused by auto-fluorescent target and effector cells and by non-specific cell death. We combined and adjusted existing cell viability assays to develop a method that allows for highly reproducible, accurate, single cell analysis by high throughput FACS, in which non-specific cell death is corrected for. In this assay the number of living, calcein AM labeled cells that are green fluorescent are quantified by adding a fixed number of unlabeled calibration beads to the analysis. Using this modified FACS calcein AM retention method, we found EC50 values to be highly reproducible and considerably lower compared to EC50 values obtained by conventional assays, displaying the high sensitivity of this assay.
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Base de dados:
MEDLINE
Assunto principal:
Células Matadoras Naturais
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Citotoxicidade Imunológica
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Citometria de Fluxo
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Fluoresceínas
Limite:
Humans
Idioma:
En
Ano de publicação:
2016
Tipo de documento:
Article