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This practical book provides the detailed methodology and expert guidance required for measuring and manipulating cytosolic ion concentrations. In addition, the strengths, weaknesses, and pitfalls of various techniques are presented. It is an invaluable source for those needing an objective evaluation of current methodologies and for those contemplating setting up such procedures. Key Features* A one-source reference for measuring and manipulating intracellular ions and for comparing and evaluating current methodologies* Includes overviews of* Optical probes and reagents* Fabrication and use of ion-selective microelectrodes* Use of NMR spectroscopy* Ionophores
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Methods in Neurosciences Volume 27 Measurement and Manipulation of Intracellular Ions Methods in Neurosciences Editor-in-Chief P. Michael Conn Methods in Neurosciences Volume 27 Measurement and Manipulation of Intracellular Ions Edited by Jacob Kraicer Department of Physiology Health Sciences Centre The University of Western Ontario London, Ontario, Canada and Human Frontier Science Program Strasbourg, France S. Jeffrey Dixon Division of Oral Biology and Department of Physiology Health Sciences Centre The University of Western Ontario London, Ontario, Canada ACADEMIC PRESS San Diego New York Boston London Sydney Tokyo Toronto Front cover photograph" Fura-2 stained neuron. Neuron from the nucleus accumbens in cell culture stained with fura-2. The image was obtained using Axon Imaging Workbench by digitizing an integrated frame 15()0 reset) from a CCD camera ICohu). The cell was illuminated at 380 nm. Courtesy of J. Plant and B. A. MacVicar, University of Calgary. This book is printed on acid-free paper. ( ~ Copyright 9 1995 by ACADEMIC PRESS, INC. All Rights Reserved. No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopy, recording, or any information storage and retrieval system, without permission in writing from the publisher. Academic Press, Inc. A Division of Harcourt Brace & Company 525 B Street, Suite 1900, San Diego, California 92101-4495 United Kinjcdrml Edition published hv Academic Press Limited 24-28 Oval Road, London NW1 7DX International Standard Serial Number: 1043-9471 International Standard Book Number: 0-12-185297-0 PRINTED IN THE UNITED STATES OF AMERICA 95 96 97 98 99 00 EB 9 8 7 6 5 4 3 2 1 To my mentors, John Logothetopoulos, Claude Fortier, and Don Hatcher JACOB KRAICER To my parents, Michael and Helen S. JEFFREY DIXON This Page Intentionally Left Blank Table of Contents Contributors to Volume 27 Preface Volumes in Series 1. Light-Optical-Based Reagents for the Measurement and Manipulation of Ions, Metabolites, and Macromolecules in Living Cells ix xiii xv 1 Kenneth A. Giuliano and D. Lansing Taylor 2. Use of Ion-Selective Microelectrodes for Recording Intracellular Ion Levels 17 Mary E. Morris 3. Use of NMR Spectroscopy to Measure Intracellular Ion Concentrations 39 Stephen J. Karlik 4. Use of Ionophores for Manipulating Intracellular Ion Concentrations 52 G. Andrew Woolley, Douglas R. Pfeiffer, and Charles M. Deber 5. Fluorescence Imaging of Cytosolic Calcium: An Introduction to Basic Experimental Principles 69 David A. Williams 6. Fluorescence Measurements of Cytosolic Calcium: Combined Photometry with Electrophysiology 81 W. T. Mason, J. Dempster, R. Zorec, John Hoyland, and P. M. Lledo 7. Use of Chelators and Photoactiva