This volume describes a range of standard and novel methodological approaches used to probe ion channel function across different modalities. Chapters guide readers through methods and protocols from an introduction to the decades old patch clamp method for the ion channel neophyte to more complex, recent protocol advances, such as optogenetics. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, application details for both the expert and non-expert reader, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Patch Clamp Electrophysiology: Methods and Protocols aims to be a reference guide for current and future ion channel physiologists.
Patch clamping is a widely applied electrophysiological technique for the study of ion channels; membrane proteins that regulate the flow of ions across cellular membranes and therefore influence the physiology of all cells. Patch Clamping aims to cover the basic principles and practical applications of this important technique. Starting with a review of the history of patch clamping, the text then goes on to cover the basic principles, platforms, equipment and environmental control, and will also include coverage of preparation types, recording modes and analysis of results. This book will explain the basic principles and practical application of patch clamp electrophysiology Written in a non-technical style to ensure its broad appeal to novice users Takes a practical approach This self-contained guide provides everything a practising patch clamp electrophysiologist needs to know to master this technique, including an overview of membrane biophysics, standard experimental design, data analysis, and technical concerns
Channels and transporters are multi-functional proteins that mediate substrate transport and signal transmission and simultaneously act as regulators for other proteins and biosensors for environmental materials. Patch clamping is an epoch-making technique that allows researchers to perform real-time measurements of electrogenic channel/transporter functions at the single/multiple molecular level. This book describes not only the conventional patch clamp techniques but also their newly developed variations or applications, such as perforated patch, slice patch, blind patch, in vivo patch, imaging patch, smart patch, and automated patch clamping. These patch clamp techniques are now essential and are extensively used across the life sciences and in related industries. With plain and practical descriptions of patch clamping and how to carry it out, especially for beginners, the book also shows how widely and exquisitely the patch clamp techniques can be applied by expert electrophysiologists. This work serves as a useful guide for young researchers and students in training and laboratory courses as well as for senior researchers who wish to extend their repertoire of techniques.
Patch-Clamp Analysis: Advanced Techniques describes in reproducible detail all applications that involve patch pipet. Beginning with updated basic patch-clamp techniques, the book presents the newest developments, ranging from fast external solution switching and the switching of the pipet solution during recording, to the loose patch, the perforated patch, and the patch cram detection technique. The advanced techniques covered combine molecular biology and imaging to produce the patch pipet with RT-PCR and fluorometric techniques.
This book is a stimulating and interesting addition to the collected works on Patch clamp technique. Patch Clamping is an electrophysiological technique, which measures the electric current generated by a living cell, due to the movement of ions through the protein channels present in the cell membrane. The technique was developed by two German scientists, Erwin Neher and Bert Sakmann, who received the Nobel Prize in 1991 in Physiology for this innovative work. Patch clamp technique is used for measuring drug effect against a series of diseases and to find out the mechanism of diseases in animals and plants. It is also most useful in finding out the structure function activities of compounds and drugs, and most leading pharmaceutical companies used this technique for their drugs before bringing them for clinical trial. This book deals with the understanding of endogenous mechanisms of cells and their receptors as well as advantages of using this technique. It covers the basic principles and preparation types and also deals with the latest developments in the traditional patch clamp technique. Some chapters in this book take the technique to a next level of modulation and novel approach. This book will be of good value for students of physiology, neuroscience, cell biology and biophysics.
Since its inception, patch-clamp has continued to be widely considered the gold standard method to record ion channel activity. Patch-Clamp Methods and Protocols, Second Edition, provides a comprehensive collection of new techniques for the development of automated, high-throughput screening systems for pharmacological evaluation, the use of various patch-clamp configurations together with novel molecular biological and imaging methodologies and enhanced stimulation protocols and perfusion systems. Divided into sections on pharmacology, physiology and biophysics, the chapters cover methods to generate more physiologically relevant conditions for drug application and screening technologies, recently developed applications such as optogenetic stimulation, advances in whole-cell recordings in freely-moving animals and novel technologies to create custom microelectrodes designed for reducing the access resistance and improving the rate of molecular diffusion. Patch-clamp is an indispensable technique for conducting pharmacological, physiological and biophysical research aimed at understanding crucial aspects of cellular and network function. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Patch-Cla mp Methods and Protocols, Second Edition will provide a useful technical and methodological guide to diverse audiences of electrophysiologists, from students to experienced investigators.
Basic Electrophysiological Methods provides a concise and easy-to-read guide on a selection of the most important contemporary electrophysiological techniques, their implementation, applications, and ways in which they can be combined and integrated with neuroscientific techniques. Intended for students, postdocs, and faculty with a basic neuroscience background, this text will not obscure the relevant technical details with textbook neuroscience tutorials as many other books do. Instead, each chapter provides a conscientious overview of the underlying theory -- a comprehensive description of equipment, materials, methods, data management, and analysis -- a troubleshooting guide, and a list of frequently asked questions. No book or online resource can function as strictly a DIY set of instructions on how to implement a complex technique. However, this book provides a fundamental and accessible set of information intended to form a foundation prior to, during, and after hands-on experience and training, greatly facilitating the initial learning process and subsequent fine-tuning of technical details.
DeFelice presents this intricate subject in an easy-to-follow, stepwise fashion: he reviews the fundamentals of electricity; transfers those principles to a biological context; and expands the discussion to encompass the subject's practical dimensions. Clear definitions and intuitive descriptions characterize the presentation, which is complemented by over 150 drawings and graphs. Mathematics is kept to the minimum necessary. The text covers both excitable and non-excitable membranes and includes the plasma membrane as well as intracellular membranes. A unique, `electronics-made-simple' appendix, designed specifically for biologists, treats the operational amplifiers used in patch clamp, and other appendices offer solutions to equations and examples that illustrate principles.
Cardiovascular disease is the major cause of mortality and morbidity in the Western Hemisphere. While significant progress has been made in treating a major sub-category of cardiac disease, arrhythmias, significant unmet needs remain. In particular, every day, thousands of patients die because of arrhythmias in the US alone, and atrial fibrillation is the most common arrhythmia affecting millions of patients in the US alone at a given time. Therefore, there is a public need to continue to develop new and better therapies for arrhythmias. Accordingly, an ever increasing number of biomedical, pharmaceutical, and medical personnel is interested in studying various aspects of arrhythmias at a basic, translational, and applied level, both in industry (ie Biotech, Pharmaceutical and device), and in academia. Not only has our overall understanding of molecular bases of disease dramatically increased, but so has the number of available and emerging molecular, pharmacological or device treatment based therapies. This practical, state-of-the art handbook will summarize and review key research methods and protocols, their advantages and pitfalls, with a focus on practical implementation, and collaborative cross-functional research. The volume will include visual and easy-to-use graphics, bulleted summaries, boxed summary paragraphs, links to reference websites, equipment manufacturers where appropriate, photographs of typical experimental setups and so forth, to keep this book very focused on practical methods and implementation, and yet, provide enough theory that the principles are clearly understood and can be easily applied.
Calcium Entry Channels in Non-Excitable Cells focuses on methods of investigating the structure and function of non-voltage gated calcium channels. Each chapter presents important discoveries in calcium entry pathways, specifically dealing with the molecular identification of store-operated calcium channels which were reviewed by earlier volumes in the Methods in Signal Transduction series. Crystallographic and pharmacological approaches to the study of calcium channels of epithelial cells are also discussed. Calcium ion is a messenger in most cell types. Whereas voltage gated calcium channels have been studied extensively, the non-voltage gated calcium entry channel genes have only been identified relatively recently. The book will fill this important niche.
