The modified FACS calcein AM retention assay: A high throughput flow cytometer based method to measure cytotoxicity: A high throughput flow cytometer based method to measure cytotoxicity

M. A. Gillissen; E. Yasuda; G. de Jong; S. E. Levie; D. Go; H. Spits; P. M. van Helden; M. D. Hazenberg

doi:10.1016/j.jim.2016.04.002

The modified FACS calcein AM retention assay: A high throughput flow cytometer based method to measure cytotoxicity: A high throughput flow cytometer based method to measure cytotoxicity

M. A. Gillissen
, E. Yasuda
, G. de Jong
, S. E. Levie
, D. Go
, H. Spits
, P. M. van Helden
, M. D. Hazenberg

Amsterdam University Medical Centers

Research output: Contribution to journal › Article › Academic › peer-review

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

Original language	English
Pages (from-to)	16-23
Number of pages	8
Journal	Journal of immunological methods
Volume	434
DOIs	https://doi.org/10.1016/j.jim.2016.04.002
Publication status	Published - 1 Jul 2016

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

ADCC
Cancer
CDC
Cellular cytotoxicity
Monoclonal antibodies

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10.1016/j.jim.2016.04.002

Cite this

Gillissen, M. A., Yasuda, E., de Jong, G., Levie, S. E., Go, D., Spits, H., van Helden, P. M., & Hazenberg, M. D. (2016). The modified FACS calcein AM retention assay: A high throughput flow cytometer based method to measure cytotoxicity: A high throughput flow cytometer based method to measure cytotoxicity. Journal of immunological methods, 434, 16-23. https://doi.org/10.1016/j.jim.2016.04.002

@article{148e2f41f9a74f7dbf92a7a6ac30f525,

title = "The modified FACS calcein AM retention assay: A high throughput flow cytometer based method to measure cytotoxicity: A high throughput flow cytometer based method to measure cytotoxicity",

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",

keywords = "ADCC, Cancer, CDC, Cellular cytotoxicity, Monoclonal antibodies",

author = "Gillissen, \{M. A.\} and E. Yasuda and \{de Jong\}, G. and Levie, \{S. E.\} and D. Go and H. Spits and \{van Helden\}, \{P. M.\} and Hazenberg, \{M. D.\}",

note = "Publisher Copyright: {\textcopyright} 2016 The Authors.",

year = "2016",

month = jul,

day = "1",

doi = "10.1016/j.jim.2016.04.002",

language = "English",

volume = "434",

pages = "16--23",

journal = "Journal of immunological methods",

issn = "0022-1759",

publisher = "Elsevier",

}

Gillissen, MA, Yasuda, E, de Jong, G, Levie, SE, Go, D, Spits, H, van Helden, PM & Hazenberg, MD 2016, 'The modified FACS calcein AM retention assay: A high throughput flow cytometer based method to measure cytotoxicity: A high throughput flow cytometer based method to measure cytotoxicity', Journal of immunological methods, vol. 434, pp. 16-23. https://doi.org/10.1016/j.jim.2016.04.002

The modified FACS calcein AM retention assay: A high throughput flow cytometer based method to measure cytotoxicity: A high throughput flow cytometer based method to measure cytotoxicity. / Gillissen, M. A.; Yasuda, E.; de Jong, G. et al.
In: Journal of immunological methods, Vol. 434, 01.07.2016, p. 16-23.

Research output: Contribution to journal › Article › Academic › peer-review

TY - JOUR

T1 - The modified FACS calcein AM retention assay: A high throughput flow cytometer based method to measure cytotoxicity

T2 - A high throughput flow cytometer based method to measure cytotoxicity

AU - Gillissen, M. A.

AU - Yasuda, E.

AU - de Jong, G.

AU - Levie, S. E.

AU - Go, D.

AU - Spits, H.

AU - van Helden, P. M.

AU - Hazenberg, M. D.

PY - 2016/7/1

Y1 - 2016/7/1

N2 - 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

AB - 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

KW - ADCC

KW - Cancer

KW - CDC

KW - Cellular cytotoxicity

KW - Monoclonal antibodies

UR - https://www.scopus.com/pages/publications/84964260958

U2 - 10.1016/j.jim.2016.04.002

DO - 10.1016/j.jim.2016.04.002

M3 - Article

C2 - 27084117

SN - 0022-1759

VL - 434

SP - 16

EP - 23

JO - Journal of immunological methods

JF - Journal of immunological methods

ER -

The modified FACS calcein AM retention assay: A high throughput flow cytometer based method to measure cytotoxicity: A high throughput flow cytometer based method to measure cytotoxicity

Abstract

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Keywords

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