Toggle Main Menu Toggle Search

Open Access padlockePrints

Measurement of Intracellular pH in Pancreatic Duct Cells: A New Method for Calibrating the Fluorescence Data

Lookup NU author(s): Dr Zoltan Rakonczay, Dr Michael Gray, Professor Barry Argent


Full text for this publication is not currently held within this repository. Alternative links are provided below where available.


Pancreatic duct cells secrete the bicarbonate ions found in pancreatic juice. Impairment of ductal bicarbonate secretion, as occurs in cystic fibrosis, has serious consequences for pancreatic function and for the structural integrity of the gland. As bicarbonate is a buffer ion, the accurate measurement of intracellular pH (pHi) in duct cells is an important technique for studying the mechanisms of bicarbonate transport. Commonly, pHi is measured using the fluorescent dye biscarboxyethylcarboxyfluorescein (BCECF). The purpose of this study was to develop a new technique for the accurate calibration of BCECF fluorescent signals. Our results indicate that BCECF fluorescence is not only dependent on pHi but also on the total fluorescence intensity of the detected area (which may be influenced by dye loading, dye leakage, and the shutter size on the photomultiplier). The outcome is that one calibration curve is not sufficient for accurate determination of pHi. In fact, an appropriate calibration curve must be selected for each individual experiment. Moreover, the calibration plot is only linear over a narrow range of pH i values. In conclusion, we have developed a new technique that should be applicable to all cell types for the accurate calibration of fluorescent signals from the pH-sensitive dye BCECF.

Publication metadata

Author(s): Hegyi P, Rakonczay Jr Z, Gray MA, Argent BE

Publication type: Article

Publication status: Published

Journal: Pancreas

Year: 2004

Volume: 28

Issue: 4

Pages: 427-434

Print publication date: 01/05/2004

ISSN (print): 0885-3177

ISSN (electronic): 1536-4828

Publisher: Lippincott Williams & Wilkins


DOI: 10.1097/00006676-200405000-00012

PubMed id: 15097861


Altmetrics provided by Altmetric