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).

Publication DOI: https://doi.org/10.1016/j.xpro.2020.100157
Divisions: College of Health & Life Sciences
College of Health & Life Sciences > School of Biosciences
College of Health & Life Sciences > School of Biosciences > Cellular and Molecular Biomedicine
Additional Information: ©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/). Funding: We acknowledge the UK Biotechnology & Biosciences Research Council (to R.M.B., A.C.C., and P.K. through BB/P025927/1); the Swedish Research Council (to S.T.-H. through 2013-05945); the Crafoord Foundation (to S.T.-H. through 20140811 and 20180916); the Magnus Bergvall Foundation (to S.T.-H. through 2015-01534); M.A.-A. is supported by a studentship co-funded by Aston University and the UK Engineering and Physical Sciences Research Council (EP/R512889/1) to R.M.B.; P.K. is the recipient of an Aston University 50th Anniversary Prize Fellowship: M.M.S. is supported through funding from Biogen to Prof. Tomas Kirchhausen. The graphical abstract was created with Biorender.com.
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Related URLs: https://www.sci ... 1441?via%3Dihub (Publisher URL)
PURE Output Type: Article
Published Date: 2020-12-18
Published Online Date: 2020-11-02
Accepted Date: 2020-11-01
Authors: Kitchen, Philip (ORCID Profile 0000-0002-1558-4673)
Salman, Mootaz M
Abir-Awan, Mohammed
Al-Jubair, Tamim
Törnroth-Horsefield, Susanna
Conner, Alex C
Bill, Roslyn M (ORCID Profile 0000-0003-1331-0852)

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