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.

Single-channel recording has become a widely used tool for the study of ion permeation mechanisms in biological membranes. Whereas the technique might have been considered an "art" after its introduction in 1976, it developed into a relatively simple method after it became possible to obtain high-resistance (several gigaohm) membrane-pipette seals. In the summer of 1982, a course on the technique was held at the Ettore Majorana Center for Scientific Culture in Erice, Sicily. It brought together people from most of the laboratories involved in patch clamping at that time. During the course, it became apparent that the technique had reached a state of maturity. Repeatedly, the opinion was expressed that a detailed description of all the aspects of the technique including representative examples of results should be available. We therefore asked the course instructors, as well as several other colleagues, to provide chapters on selected topics in order to produce this volume. The different variants of patch clamping were described quite extensively in an article by Hamill, Marty, Neher, Sakmann, and Sigworth (Pflugers Archiv 391:85) in 1981. Rather than repeating this survey in an introductory chapter, we chose to reprint that article in the Appendix of this volume (by permission of Springer-Verlag). The methods section will, therefore, go straight into detailed aspects of the technol ogy.

The propagation of signals through the nervous system depends on rapid changes in electric potential across cell membranes. These changes are mediated by ion channels--macromolecular pores that facilitate the passage of specific ions (e.g., K+ or Na+) through cell membranes in response to various signals. Defects in ion channels can lead to diseases such as epilepsy. This laboratory manual provides state-of-the-art techniques for investigating ion channel properties and activity, particularly in the nervous system. Contributors present electrophysiological methods to examine single-channel activity in cultured cells, to study synaptic plasticity and circuit dynamics in brain slice preparations, and to perform whole-cell recordings in awake--and even freely moving--animals. The use of optogenetic tools to study synapses or small networks in organotypic slice cultures is also covered. Many of the experimental setups described can be adapted for other ion channels, cell types, or systems. The manual includes background on the structure, function, and regulation of different voltage- and ligand-gated ion channels. Therefore, it is a useful resource for all cell biologists and neuroscientists seeking to further understand the complex roles of ion channels in normal physiology and disease.

Despite the many milestones in cystic fibrosis (CF) research, progress towards curing the disease has been slow, and it is increasingly difficult to grasp and use the already wide and still growing range of diverse methods currently employed to study CF so as to understand it in its multidisciplinary nature. Cystic Fibrosis: Diagnosis and Protocols aims to provide the CF research community and related researchers with a very wide range of high-quality experimental tools, as an easy way to grasp and use classical and novel methods applied to cystic fibrosis. Volume I: Approaches to Study and Correct CFTR Defects focuses on the cystic fibrosis transmembrane conductance regulator (CFTR) and its expression, biogenesis, structure, and function in terms of the defects causing CF. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and practical, Cystic Fibrosis: Diagnosis and Protocols will provide readers with optimal working tools to address pressing questions in the best technical way, while helping all of us, as a research and clinical community, to move faster hand-in-hand toward unravelling the secrets of this challenging disorder and cure it.

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.

The New Benchmark for Understanding the Latest Developments of Ion ChannelsIon channels control the electrical properties of neurons and cardiac cells, mediate the detection and response to sensory stimuli, and regulate the response to physical stimuli. They can often interact with the cellular environment due to their location at the surface of ce

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.