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An improved flow cytometry assay to monitor phagosome acidification

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An improved flow cytometry assay to monitor phagosome acidification. / Colas, Chloé; Menezes, Shinelle; Gutiérrez-Martínez, Enric; Péan, Claire B.; Dionne, Marc S.; Guermonprez, Pierre.

In: Journal of Immunological Methods, Vol. 412, 01.10.2014, p. 1-13.

Research output: Contribution to journalArticle

Harvard

Colas, C, Menezes, S, Gutiérrez-Martínez, E, Péan, CB, Dionne, MS & Guermonprez, P 2014, 'An improved flow cytometry assay to monitor phagosome acidification', Journal of Immunological Methods, vol. 412, pp. 1-13. https://doi.org/10.1016/j.jim.2014.06.008

APA

Colas, C., Menezes, S., Gutiérrez-Martínez, E., Péan, C. B., Dionne, M. S., & Guermonprez, P. (2014). An improved flow cytometry assay to monitor phagosome acidification. Journal of Immunological Methods, 412, 1-13. https://doi.org/10.1016/j.jim.2014.06.008

Vancouver

Colas C, Menezes S, Gutiérrez-Martínez E, Péan CB, Dionne MS, Guermonprez P. An improved flow cytometry assay to monitor phagosome acidification. Journal of Immunological Methods. 2014 Oct 1;412:1-13. https://doi.org/10.1016/j.jim.2014.06.008

Author

Colas, Chloé ; Menezes, Shinelle ; Gutiérrez-Martínez, Enric ; Péan, Claire B. ; Dionne, Marc S. ; Guermonprez, Pierre. / An improved flow cytometry assay to monitor phagosome acidification. In: Journal of Immunological Methods. 2014 ; Vol. 412. pp. 1-13.

Bibtex Download

@article{e4caf7bd1a7b4f1faea5dcf8fc553ca4,
title = "An improved flow cytometry assay to monitor phagosome acidification",
abstract = "Phago-lysosome formation is important for cell-autonomous immunity to intracellular pathogens, antigen presentation and metabolism. A hallmark feature of phago-lysosomal compartments is that they undergo progressive luminal acidification controlled by the activation of vacuolar V-ATPase. Acidification is required for many enzymatic processes taking place in phago-lysosomes, like proteolysis, and supports the microbicidal activity of macrophages. Here we present a new quantitative methodology to assess phagosome acidification by flow cytometry based on the use of bi-fluorescent particles. This method relies on the use of UV polystyrene beads labelled with the acid sensor pHrodo-succinimidyl ester (pHrodoTM SE red) and enables us to dissociate particle association with phagocytes from their engulfment in acidified compartments. This methodology is well suited to monitor the acidification of phagosomes formed in vivo after fluorescent bead administration.",
keywords = "Flow cytometry, Lysosome, Phagocyte, Phagosome",
author = "Chlo{\'e} Colas and Shinelle Menezes and Enric Guti{\'e}rrez-Mart{\'i}nez and P{\'e}an, {Claire B.} and Dionne, {Marc S.} and Pierre Guermonprez",
year = "2014",
month = oct,
day = "1",
doi = "10.1016/j.jim.2014.06.008",
language = "English",
volume = "412",
pages = "1--13",
journal = "Journal of Immunological Methods",
issn = "0022-1759",

}

RIS (suitable for import to EndNote) Download

TY - JOUR

T1 - An improved flow cytometry assay to monitor phagosome acidification

AU - Colas, Chloé

AU - Menezes, Shinelle

AU - Gutiérrez-Martínez, Enric

AU - Péan, Claire B.

AU - Dionne, Marc S.

AU - Guermonprez, Pierre

PY - 2014/10/1

Y1 - 2014/10/1

N2 - Phago-lysosome formation is important for cell-autonomous immunity to intracellular pathogens, antigen presentation and metabolism. A hallmark feature of phago-lysosomal compartments is that they undergo progressive luminal acidification controlled by the activation of vacuolar V-ATPase. Acidification is required for many enzymatic processes taking place in phago-lysosomes, like proteolysis, and supports the microbicidal activity of macrophages. Here we present a new quantitative methodology to assess phagosome acidification by flow cytometry based on the use of bi-fluorescent particles. This method relies on the use of UV polystyrene beads labelled with the acid sensor pHrodo-succinimidyl ester (pHrodoTM SE red) and enables us to dissociate particle association with phagocytes from their engulfment in acidified compartments. This methodology is well suited to monitor the acidification of phagosomes formed in vivo after fluorescent bead administration.

AB - Phago-lysosome formation is important for cell-autonomous immunity to intracellular pathogens, antigen presentation and metabolism. A hallmark feature of phago-lysosomal compartments is that they undergo progressive luminal acidification controlled by the activation of vacuolar V-ATPase. Acidification is required for many enzymatic processes taking place in phago-lysosomes, like proteolysis, and supports the microbicidal activity of macrophages. Here we present a new quantitative methodology to assess phagosome acidification by flow cytometry based on the use of bi-fluorescent particles. This method relies on the use of UV polystyrene beads labelled with the acid sensor pHrodo-succinimidyl ester (pHrodoTM SE red) and enables us to dissociate particle association with phagocytes from their engulfment in acidified compartments. This methodology is well suited to monitor the acidification of phagosomes formed in vivo after fluorescent bead administration.

KW - Flow cytometry

KW - Lysosome

KW - Phagocyte

KW - Phagosome

UR - http://www.scopus.com/inward/record.url?scp=84908306680&partnerID=8YFLogxK

U2 - 10.1016/j.jim.2014.06.008

DO - 10.1016/j.jim.2014.06.008

M3 - Article

AN - SCOPUS:84908306680

VL - 412

SP - 1

EP - 13

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

SN - 0022-1759

ER -

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