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The Axon Guide for Electrophysiology & Biophysics Laboratory Techniques Axon Instruments, Inc. Part Number 2500-102 Rev A Printed in U.S.A. PREFACE Axon Instruments, Inc. is pleased to present you with The Axon Guide, a laboratory guide to electrophysiology and biophysics. The purpose of this Guide is to serve as a state-of-the-art information and data resource. It covers a broad scope of topics ranging from the biological basis of bioelectricity and a description of the basic experimental setup to the principles of operation of the most advanced hardware and software equipment currently available. We planned the Guide as a tool benefiting both the novice and the expert electrophysiologist. Newcomers to electrophysiology will gain an appreciation of the intricacies of electrophysiological measurements and the requirements for setting up a complete recording and analysis system. For experienced electrophysiologists, we included in-depth discussions of selected topics of hardware and software design and their applications. We undertook this endeavor with great enthusiasm. We believe that The Axon Guide is a valuable contribution to our customers and to the field of electrophysiology and biophysics. We are planning to update this Guide periodically. However, the success of this Guide in achieving its goals depends on your input. We encourage you to contact us with comments on the Guide's content, style and usefulness to you and to your laboratory. Your comments will assist us in improving the next edition of the Guide. If you would like to contribute to a future edition, please contact us. This Guide is a product of a collaborative effort of researchers active in the field of electrophysiology and of Axon Instruments' staff. The names of the authors along with the editorial staff are listed on the following pages. I am deeply grateful to these individuals for sharing their knowledge and devoting significant time and effort to this endeavor. I would also like to thank Dr. Alan Finkel, the founder and CEO of Axon Instruments, and Geoff Powell, President of Axon Instruments, for their support and encouragement. The editorial assistance, layout and graphics done expertly by Jay Kurtz is much appreciated. Many thanks to Axon Instruments' personnel for assisting in numerous ways. Rivka Sherman-Gold, Ph.D. Editor, The Axon Guide Director, Marketing & Technical Support Axon Instruments, Inc. June 1993 INTRODUCTION Axon Instruments, Inc., was founded in 1983 to design and manufacture instrumentation and software for electrophysiology and biophysics. Our products are used primarily in cellular neuroscience and cardiovascular research. Most applications are at the cellular and molecular level, such as searching for potent Ca-channel blockers, elucidating the molecular mechanisms involved in learning and memory and investigating the control mechanisms of vasodilation and constriction. However, the applications for our products are not limited to traditional nervous system research. They include general physiology studies at the isolated-organ and whole-animal level (e.g., pulmonary, auditory, and vascular research), and plant electrophysiology. Axon Instruments products are distinguished by their innovative design, high quality manufacturing standards and expert technical support. Our goal is to respond to the needs of researchers by providing them with the best possible equipment and applications information. Our growth over the past ten years has been paralleled by a period of rapid growth in neuroscience research. Despite the fact that total research funding has increased significantly, so too have the number of researchers competing for these funds. The maturation of the field and the tight competition for funds has encouraged an ever increasing percentage of researchers to turn to commercial sources for equipment and software,. instead of trying to personally develop the required research tools. Axon Instruments has responded to this demand by providing ready- to-use, technologically advanced instrumentation and software, thereby enabling researchers to spend their time in the pursuit of their primary research goals. Based on the feedback that we receive from our customers, we continually enhance our products so that we can keep abreast and ahead of researchers' needs. To help our customers derive maximal advantage from our products, we have, from our inception, given priority to our technical support department, whose staff includes experienced electrophysiologists. As science is becoming more integrated in the approaches researchers use, and more molecular biologists, biochemists and pharmacologists employ electrophysiological techniques, Axon Instruments is increasingly assuming a significant role in guiding newcomers into the field and disseminating information about technological advancements. In addition to describing the operation of products, our product manuals include relevant background information and related experimental techniques. Our newsletter, AxoBits, serves as a forum to disseminate information on our products as well as on the latest experimental techniques employed by our customers. And finally, for the first time, we are publishing a laboratory techniques workbook, The Axon Guide. Page iv The 1993 Axon Guide is the first publication of this kind that we have produced. The Axon Guide would never have been completed without the superb effort of Dr. Rivka Sherman-Gold, the Guide's editor. I wish to congratulate Rivka for the skill and dedication that she brought to this task. Alan S. Finkel, Ph.D. Chief Executive Officer Axon Instruments, Inc. June 1993 Page v Acknowledgment to Axon Consultants and Customers Axon Instruments employs a talented team of engineers and scientists dedicated to designing instruments and software incorporating the most advanced technology and the highest quality. Nevertheless, it would not be possible for us to enhance our products without close collaborations with members of the scientific community. These collaborations take many forms. Some scientists assist Axon Instruments on a regular basis, sharing their insights on current needs and future directions of scientific research. Others assist us by virtue of a direct collaboration in the design of certain products. Many scientists help us by reviewing our instrument designs and the development versions of various software products. We are grateful to these scientists for their assistance. We also receive a significant number of excellent suggestions from the customers we meet at scientific conferences. To all of you who have visited us at our booths and shared your thoughts, we extend our sincere thanks. Another source of feedback for us is the information that we receive from the conveners of the many excellent summer courses and workshops that we support with equipment loans. Our gratitude is extended to them for the written assessments they often send us outlining the strengths and weaknesses of the Axon Instruments products. AXON GUIDE Page vi Company Philosophy Axon Instruments is a technology-driven company. That is, our sales growth is expected to derive from the introduction of technologically advanced products. These products are expected to embody superb elegance, value and functionality. We are responsive to the suggestions and criticisms made by our customers. These ideas and comments guide us in the continued enhancement of our existing products and the design of new products. We recognize our customers' need for prompt and courteous service. This means that we provide the highest-caliber technical support, fast repair service, responsive order processing and sales support, and polite attention on the phone and at conferences. We sell our products direct around the world in the belief that direct contact with our customers helps us to provide them with the best possible service and support. We are proud of our reputation as the world's best manufacturer of instruments for electrophysiological research. We continually strive to maintain this reputation through quality of design, manufacture, product testing and technical support. Page vii pCLAMP Users' Group The pCLAMP Users' Group was launched in April 1990 with the release of the first issue of the group's newsletter, The Channel. The newsletter and the users' group was founded by an independent researcher whose aim was to provide a forum for the use of pCLAMP that was independent of Axon Instruments, Inc. Although Axon Instruments, Inc. underwrites the costs of printing and mailing the PUG newsletter, its editorial contents are independent of Axon Instruments. Newsletters are mailed at irregular intervals to all registered users. New users may receive back issue of the newsletter and a pCLAMP Users' Group Director by writing to: The pCLAMP Users' Group 725 Huckins Road Cheshire, CT 06410 USA AXON GUIDE Page viii Customer Feedback We urge you to write to us if you have any suggestions for bettering our products or our customer service. Your letters should be addressed to the Product Development Manager or the Customer Service Manager at Axon Instruments, Inc., 1101 Chess Drive, Foster City, CA 94404, USA. Fax: +1(650)571-9500 We would appreciate hearing from you, whether you are the original purchaser, an inheritor, a student user, or if you purchased the product from a distributor. Many of you have ideas for new products. We are always interested in discussing the possibility of collaboration with our customers. Please contact us if you have an idea for a new product that you think might have commercial significance. It is better to contact us early so that we can evaluate the commercial potential of your ideas before you have expended too much effort and also so that we can participate in the design process as early as possible. CONTRIBUTORS The Axon Guide is the product of a collaborative effort of Axon Instruments, Inc. and researchers from several institutions whose contributions to the Guide are gratefully acknowledged. John M. Bekkers, Ph.D. Division of Neuroscience, John Curtin School of Medical Research, Australian National University, Canberra, A.C.T. Australia Richard J. Bookman, Ph.D. Department of Pharmacology, School of Medicine, University of Miami, Miami, Florida Michael J. Delay, Ph.D. Axon Instruments, Inc., Foster City, California Alan S. Finkel, Ph.D. Axon Instruments, Inc., Foster City, California Aaron P. Fox, Ph.D. Department of Pharmacological & Physiological Sciences, University of Chicago, Chicago, Illinois David Gallegos Axon Instruments, Inc., Foster City, California Robert I. Griffiths, Ph.D. Centre for Early Human Development, Medical Centre, Monash University, Clayton, Victoria, Australia Donald Hilgemann, Ph.D. Department of Physiology, University of Texas, Dallas, Texas Richard H. Kramer, Ph.D. Center for Neurobiology and Behavior, Columbia Univeristy, College of Physicians and Surgeons, New Yortk, NY Henry A. Lester, Ph.D. Division of Biology, California Institute of Technology, Pasadena, California Richard A. Levis, Ph.D. Department of Physiology, Rush-Presbyterian-St. Luke's Medical College, Chicago, Illinois Edwin S. Levitan, Ph.D. Department of Pharmacology, School of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania M. Craig McKay, Ph.D. Bristol-Myers Squibb, Pharmaceutical Research Institute Department of Biophysics & Molecular Biology, Wallingford, CT David J. Perkel, Ph.D. Department of Pharmacology, School of Medicine, University of California, San Francisco, San Francisco, California Stuart H. Thompson, Ph.D. Hopkins Marine Station, Stanford University, Pacific Grove, California James L. Rae, Ph.D. Department of Physiology and Biophysics, Mayo Clinic, Rochester, Minnesota Michael M. White, Ph.D. Department of Physiology, Medical College of Pennsylvania, Philadelphia, Pennsylvania William F. Wonderlin, Ph.D. Department of Phamacology and Toxicology, Health Science Center, West Virginia University, Morgantown, West Virginia AXON GUIDE EDITORIAL Editor Rivka Sherman-Gold, Ph.D. Editorial Committee Alan S. Finkel, Ph.D Henry A. Lester, Ph.D* Michael J. Delay, Ph.D Rivka Sherman-Gold, Ph.D W. Geoff Powell Assistant Editor Jay Kurtz Artwork Elizabeth Brzeski * H. A. Lester is at the California Institute of Technology, Pasadena, California. The other editorial staff are at Axon Instruments. Acknowledgements The valuable inputs and insightful comments of Drs. Bertil Hille, Joe Immel and Stephen Redman and Mr. Burt Maertz are much appreciated.