Calcein Fluorescence Quenching to Measure Plasma Membrane Water Flux in Live Mammalian Cells

Philip Kitchen; Mootaz M Salman; Mohammed Abir-Awan; Tamim Al-Jubair; Susanna Törnroth-Horsefield; Alex C Conner; Roslyn M Bill

doi:10.1016/j.xpro.2020.100157

Calcein Fluorescence Quenching to Measure Plasma Membrane Water Flux in Live Mammalian Cells

Philip Kitchen, Mootaz M Salman, Mohammed Abir-Awan, Tamim Al-Jubair, Susanna Törnroth-Horsefield, Alex C Conner, Roslyn M Bill

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

Abstract

Aquaporins (AQPs) are membrane channel proteins that facilitate the movement of water down osmotic gradients across biological membranes. This protocol allows measurements of AQP-mediated water transport across the plasma membrane of live mammalian cells. Calcein is a fluorescent dye that is quenched in a concentration-dependent manner. Therefore, on short timescales, its concentration-dependent fluorescence can be used as a probe of cell volume, and therefore a probe of water transport into or out of cells. For complete details on the use and execution of this protocol, please refer to Kitchen et al. (2020) and Kitchen and Conner (2015). For the underlying methodology development, please refer to Fenton et al. (2010) and Solenov et al. (2004).

Original language	English
Article number	100157
Journal	STAR protocols
Volume	1
Issue number	3
Early online date	2 Nov 2020
DOIs	https://doi.org/10.1016/j.xpro.2020.100157
Publication status	Published - 18 Dec 2020

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10.1016/j.xpro.2020.100157Licence: CC BY-NC-ND

Calcein Fluorescence Quenching
©2020 The Author(s). This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).1llOPENACCESS
Final published version, 1.55 MBLicence: CC BY-NC-ND

https://www.sciencedirect.com/science/article/pii/S2666166720301441?via%3Dihub

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title = "Calcein Fluorescence Quenching to Measure Plasma Membrane Water Flux in Live Mammalian Cells",

abstract = "Aquaporins (AQPs) are membrane channel proteins that facilitate the movement of water down osmotic gradients across biological membranes. This protocol allows measurements of AQP-mediated water transport across the plasma membrane of live mammalian cells. Calcein is a fluorescent dye that is quenched in a concentration-dependent manner. Therefore, on short timescales, its concentration-dependent fluorescence can be used as a probe of cell volume, and therefore a probe of water transport into or out of cells. For complete details on the use and execution of this protocol, please refer to Kitchen et al. (2020) and Kitchen and Conner (2015). For the underlying methodology development, please refer to Fenton et al. (2010) and Solenov et al. (2004).",

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AU - Kitchen, Philip

AU - Salman, Mootaz M

AU - Abir-Awan, Mohammed

AU - Al-Jubair, Tamim

AU - Törnroth-Horsefield, Susanna

AU - Conner, Alex C

AU - Bill, Roslyn M

PY - 2020/12/18

Y1 - 2020/12/18

N2 - Aquaporins (AQPs) are membrane channel proteins that facilitate the movement of water down osmotic gradients across biological membranes. This protocol allows measurements of AQP-mediated water transport across the plasma membrane of live mammalian cells. Calcein is a fluorescent dye that is quenched in a concentration-dependent manner. Therefore, on short timescales, its concentration-dependent fluorescence can be used as a probe of cell volume, and therefore a probe of water transport into or out of cells. For complete details on the use and execution of this protocol, please refer to Kitchen et al. (2020) and Kitchen and Conner (2015). For the underlying methodology development, please refer to Fenton et al. (2010) and Solenov et al. (2004).

AB - Aquaporins (AQPs) are membrane channel proteins that facilitate the movement of water down osmotic gradients across biological membranes. This protocol allows measurements of AQP-mediated water transport across the plasma membrane of live mammalian cells. Calcein is a fluorescent dye that is quenched in a concentration-dependent manner. Therefore, on short timescales, its concentration-dependent fluorescence can be used as a probe of cell volume, and therefore a probe of water transport into or out of cells. For complete details on the use and execution of this protocol, please refer to Kitchen et al. (2020) and Kitchen and Conner (2015). For the underlying methodology development, please refer to Fenton et al. (2010) and Solenov et al. (2004).

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