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J Cyst Fibros. 2004 Aug;3 Suppl 2:171-6.

Measurement of halide efflux from cultured and primary airway epithelial cells using fluorescence indicators.

Author information

1
Department of Gene Therapy, National Heart and Lung Institute, Faculty of Medicine, Imperial College London, London SW3 GLR, UK.

Abstract

The use of the halide-sensitive fluorescent probes (6-methoxy-N-(-sulphopropyl)quinolinium (SPQ) and N-(ethoxycarbonylmethyl)-6-methoxyquinolinium bromide (MQAE)) to measure chloride transport in cells has now been established as an alternative to the halide-selective electrode technique, radioisotope efflux assays and patch-clamp electrophysiology. We report here procedures for the assessment of halide efflux, using SPQ/MQAE halide-sensitive fluorescent indicators, from both adherent cultured epithelial cells and freshly obtained primary human airway epithelial cells. The procedure describes the calculation of efflux rate constants using experimentally derived SPQ/MQAE fluorescence intensities and empirically derived Stern-Volmer calibration constants. These fluorescence methods permit the quantitative analysis of CFTR function.

PMID:
15463953
DOI:
10.1016/j.jcf.2004.05.036
[Indexed for MEDLINE]
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