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

Felix Munkonge, Eric W F W Alton, Charlotte Andersson, Heather Davidson, Anca Dragomir, Aleksander Edelman, Ray Farley, Lena Hjelte, Gerry McLachlan, Myra Stern, Godfried M Roomans

Research output: Contribution to journalArticlepeer-review

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.
Original languageEnglish
Pages (from-to)171-6
Number of pages6
JournalJournal of Cystic Fibrosis
Volume3 Suppl 2
DOIs
Publication statusPublished - 2004

Keywords

  • Chloride transport;
  • Airway epithelial cell
  • SPQ
  • MQAE
  • FLUORESCENCE MICROSCOPY
  • CFTR

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