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      Intracellular pH Measurement with Fluorescent Dye in Canine Basilar Arteries

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          Abstract

          The intracellular pH (pH<sub>i</sub>) of basilar artery rings was determined with 2’,7’-bis-(carboxyethyl)-5,(6)-carboxyfluorescein (BCECF) by measuring the ratio of emitted (540 nm) fluorescence intensities (FI) at excitation wavelengths of 500 and 440 nm. There was a dye loss from the rings in 90 min (39.3 ± 3.6%; p < 0.001). We found that the ratio of fluorescence intensities does not adequately correct for dye loss; hence, we derived a method to correct for dye loss during pHi determinations. Calibration curves of the ratio versus pH, were constructed for the artery rings. The slope and intercept of the calibration curves depended on FI<sub>440</sub>. Linear regression lines for the slope and intercept versus FI440 were: slope = 0.00343 × FI<sub>440</sub> + 0.32484 (r – 0.96; p < 0.0001) and intercept = –0.02048 × FI<sub>440</sub> – 0.80123 (r = –0.97; p < 0.0001). In solutions with different pH and different concentrations of free BCECF, the slope of the ratio versus pH of the solution was steeper at high concentrations of BCECF. Thus, pH was calculated from a calibration curve in which the slope and intercept were determined from FI440 with the above formula. The corrected pH<sub>i</sub> was 7.37 ± 0.05 (n = 25) at pH<sub>o</sub> 7.4 and 37 °C.

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          Author and article information

          Journal
          JVR
          J Vasc Res
          10.1159/issn.1018-1172
          Journal of Vascular Research
          S. Karger AG
          1018-1172
          1423-0135
          1991
          1991
          23 September 2008
          : 28
          : 6
          : 464-474
          Affiliations
          Departments of aPharmacology and bAnesthesia, University of Manitoba, Winnipeg, Canada
          Article
          158893 Blood Vessels 1991;28:464–474
          10.1159/000158893
          © 1991 S. Karger AG, Basel

          Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher. Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug. Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

          Page count
          Pages: 11
          Categories
          Research Paper

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