cell culture methods in neurosciences

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Methods In Neurosciences

Author : P. Michael Conn
ISBN : 0121852679
Genre :
File Size : 84. 92 MB
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Cell Culture In The Neurosciences

Author : Jane Bottenstein
ISBN : 9781461324737
Genre : Medical
File Size : 52. 46 MB
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A fundamental problem in neuroscience is the elucidation of the cellular and molecular mechanisms underlying the development and function of the nervous system. The complexity of organization, the heteroge neity of cell types and their interactions, and the difficulty of controlling experimental variables in intact organisms make this a formidable task. Because of the ability that it affords to analyze smaller components of the nervous system (even single cells in some cases) and to better control experimental variables, cell culture has become an increasingly valuable tool for neuroscientists. Many aspects of neural development, such as proliferation, differentiation, synaptogenesis, and myelination, occur in culture with time courses remarkably similar to those in vivo. Thus, in vitro methods often provide excellent model systems for investigating neurobiological questions. Ross Harrison described the first culture of neural tissue in 1907 and used morphological methods to analyze the cultures. Since that time the technique has been progressively modified and used to address an ever widening range of developmental questions. In recent years a con vergence of new or improved cell culture, biochemical, electrophysiol ogical, and immunological methods has occurred and been brought to bear on neurobiological questions. This volume is intended not to be comprehensive but rather to highlight some of the latest findings, with a review of previous important work as well, in which combinations of these methods are used.

Electrochemical Methods For Neuroscience

Author : Adrian C. Michael
ISBN : 9781420005868
Genre : Science
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Since the first implant of a carbon microelectrode in a rat 35 years ago, there have been substantial advances in the sensitivity, selectivity and temporal resolution of electrochemical techniques. Today, these methods provide neurochemical information that is not accessible by other means. The growing recognition of the versatility of electrochemical techniques indicates a need for a greater understanding of the scientific foundation and use of these powerful tools. Electrochemical Methods for Neuroscience provides an updated summary of the current, albeit evolving, state of the art and lays the scientific foundation for incorporating electrochemical techniques into on-going or newly emerging research programs in the neuroscience disciplines. With contributions from pioneers in the field, the text outlines the applications and benefits of a wide range of electrochemical techniques. It explores the methodology behind the acquisition of neurochemical and neurobiological data through continuous amperometry, fast scan cyclic voltammetry, high-speed chronoamperometry, ion-selective microelectrodes, enzyme based microelectrodes, and in vivo voltammetry with telemetry. The text also introduces emerging concepts in the field such as the correlation of electrochemical recordings with information obtained from patch clamp, electrophysiological, and behavioral techniques. By presenting up-to-date information on the growing collection of electrochemical methods, microsensors, and research techniques, Electrochemical Methods for Neuroscience assists seasoned researchers and newcomers to the field in making sound decisions about adopting the most appropriate of these tools for their future research objectives.

Cellular And Molecular Methods In Neuroscience Research

Author : Adalberto Merighi
ISBN : 9780387224602
Genre : Medical
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There are numerous books on cellular and molecular protocols for general use in cell biology but very few are exclusively devoted to neurobiology. This book fills this gap and explains in a clear and consistent manner, some of the more commonly used protocols in neuroscience research. Each chapter is written by either the person who invented the procedure or an expert in the field. The format is uniform: "Overview," "Background," "Protocols," and "results and discussion." Each protocol begins with the principle of the technique, studies in cell culture, materials and reagents, and, lastly, step-by-step outline of the procedure itself. This highly practical book is also well illustrated (with 17 four color plates) to make the concepts and procedures easy to understand and perform.

Tissue Culture Techniques

Author : Bernice M. Martin
ISBN : 0817636439
Genre : Medical
File Size : 34. 5 MB
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ACKNOWLEDGMENTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Xl II INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . I STERILITY . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 Aseptic Technique . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 Physical manipulations • Use of the sterile cabinet (hood) Sterilization Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 Heat • Radiation • Toxic gas • Filtration • Antibiotics Quality Control of Sterilization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23 Routine labeling Suggested Readings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25 Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26 vi CONTENTS ROUTINE CELL CULTURE . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 Feeding Schedules and Media Components . . . . . . . . . . . . . . . . . . . . 29 General properties of media and salt solutions • water as a reagent· Establishingfeeding schedules Subcultivation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 Solutions and methods for adherent cells • Common enzyme solutions • Inoculating (seeding) the cultures Cell Enumeration and Cell Viability . . . . . . . . . . . . . . . . . . . . . . . . . . 54 Hemocytometer • Particle counter • Cell viability Putting Routine Methods to Work . . . . . . . . . . . . . . . . . . . . . . . . . . . 63 Normal cell growth characteristics Detecting and Disposing of Contamination . . . . . . . . . . . . . . . . . . . . 66 Bacteria and fUngi • Fungi • Mycoplasma • Viruses • Dealing with contamination Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73 Inadequate cell growth • Recurrent contamination • When to call your vendor Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80 Biological hazards • Chemical hazards Suggested Readings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85 Problem Set . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85 Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89 EXPERIMENTS IN CULTURE . . . . . . . . . . . . . . . . . . . . . . . . . . 91 II Alterations of the Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91 Serum • Treatments of serum • Plasma-derived serum • Serum-free and low-protein media Substrata. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 101 Coatingplasticware with solutions • Alterations with polymers • Using cells to coat the plasticware • Culturing cells on microcarriers Altering the Environment. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 106 Temperature changes • Gaseous changes Problem Set . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110 Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110 CONTENTS vii PRIMARY CELL CULTURE. . . . . . . . . . . . . . . . . . . . . . . . . . . . 113 Isolation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114 Dissection • Enzymatic dissociation methods • Nonenzymatic isolation • Purification of cell suspensions • Consideringyield and survival Chatacterization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Methods In Neurobiology

Author : Robert Lahue
ISBN : 9781468438062
Genre : Medical
File Size : 25. 11 MB
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Rapid advances in knowledge have led to an increasing interest in neuro biology over the last several years. These advances have been made possible, at least in part, by the use of increasingly sophisticated methodology. Furthermore, research in the most rapidly advancing areas is essentially multidisciplinary and is characterized by contributions from many investi gators employing a variety of techniques. While a grasp of fundamental neurobiological concepts is an obvious prerequisite for those who wish to follow or participate in this field, critical awareness and evaluation of neurobiological research also requires an understanding of sophisticated methodologies. The objective of Methods in Neurobiology is the development of such critical abilities. The reader is exposed to the basic concepts, principles, and instrumentation of key methodologies, and the application of each meth odology is placed in the special context of neurobiological research. The reader will gain familiarity with the terminology and procedures of each method and the ability to evaluate results in light of the particular features of neurobiological preparations and applications.

Protocols For Neural Cell Culture

Author : Sergey Fedoroff
ISBN : 9780896039025
Genre : Medical
File Size : 67. 80 MB
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The third edition of a collection of reproducible methods used in the neuroscience laboratory. The chapters carried over from the second edition have been extensively revised and updated, while 11 new chapters have been added which come from well-established tissue culture laboratories with proven records. The new chapters deal with preparation of substrata, use of serum-free media, maintaining hybridomas, and the production and purification of monoclonal antibodies. Also included are new and extensive chapters on the isolation, growth, and characterization of neural stem cells and glia progenitor cells.

Modern Techniques In Neuroscience Research

Author : Uwe Windhorst
ISBN : 9783642585524
Genre : Medical
File Size : 30. 12 MB
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An overview of the techniques used in modern neuroscience research with the emphasis on showing how different techniques can optimally be combined in the study of problems that arise at some levels of nervous system organization. This is essentially a working tool for the scientist in the laboratory and clinic, providing detailed step-by-step protocols with tips and recommendations. Most chapters and protocols are organized such that they can be used independently, while cross-references between the chapters, a glossary, a list of suppliers and appendices provide further help.

Practical Cell Culture Techniques

Author : Alan A. Boulton
ISBN : 0896033481
Genre : Medical
File Size : 21. 49 MB
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Most cells will survive removal from the natural mic- environment of their in vivo tissue and placement into a sterile culture dish under optimal conditions. Not only do they survive, but they also multiply and express differen- ated properties in such a culture dish. A few cells do this in suspension, but most will need some kind of mechanical support substituting for their natural connections with other cells. The surface of a culture dish that might have to be coated is usually sufficient. The recent trend to standa- ization of conditions and the existence of commercial ent- prises with adequate funds and specializing in the needs of scientists were responsible for the tremendous proliferation of cell culture techniques in all fields of research in the last 20 years. No longer does a scientist have to concentrate all his/her efforts on that technology; the new trends make it feasible to employ cell culture techniques as only one of the many methods available in a small corner of a larger research laboratory. Some areas of research depend more heavily than others on cell culture techniques. Neuroscience is one of the areas that has developed hand in hand with the prol- eration of cell culture methodology. Molecular biological aspects, cell differentiation and development, neurophy- ological and neurochemical studies, as well as investigations into the nature of various diseases are now to a large extent dependent on the use of cell cultures.

Guide To Research Techniques In Neuroscience

Author : Matt Carter
ISBN : 9780128005972
Genre : Science
File Size : 31. 30 MB
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Neuroscience is, by definition, a multidisciplinary field: some scientists study genes and proteins at the molecular level while others study neural circuitry using electrophysiology and high-resolution optics. A single topic can be studied using techniques from genetics, imaging, biochemistry, or electrophysiology. Therefore, it can be daunting for young scientists or anyone new to neuroscience to learn how to read the primary literature and develop their own experiments. This volume addresses that gap, gathering multidisciplinary knowledge and providing tools for understanding the neuroscience techniques that are essential to the field, and allowing the reader to design experiments in a variety of neuroscience disciplines. Written to provide a "hands-on" approach for graduate students, postdocs, or anyone new to the neurosciences Techniques within one field are compared, allowing readers to select the best techniques for their own work Includes key articles, books, and protocols for additional detailed study Data analysis boxes in each chapter help with data interpretation and offer guidelines on how best to represent results Walk-through boxes guide readers step-by-step through experiments

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