This manual describes the methodology and applications of microinjection, a powerful technique for transposing material into living cells through needle injection. Information is presented on the techniques principles, the required equipment, preparation of receiving cells and injected material, and the analysis of results. Detailed protocols for a wide range of applications in different cellular systems are included. A large part of the book is also dedicated to the Xenopus laevis oocyte system.

This detailed book explores how microinjection will be used in the foreseeable future, not only for generating animal models for biomedical research but also for changing economically or ecologically important species that can broadly impact our society in general. The opening half of the book focuses on methods for generating mouse models, as they are still the most popular in genome engineering research, while the second half examines gene-editing in a variety of other species, opened up by the developments in ZFN, TALEN, and CRISPR techniques. Written for the highly successful Methods in Molecular Biology series, 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. Authoritative and practical, Microinjection: Methods and Protocols serves as an ideal guide for researchers looking to take advantage of the breakthrough technologies in gene-editing and embryo micromanipulations.

This manual describes the methodology and applications of microinjection, a powerful technique for transposing material into living cells through needle injection. Information is presented on the techniques principles, the required equipment, preparation of receiving cells and injected material, and the analysis of results. Detailed protocols for a wide range of applications in different cellular systems are included. A large part of the book is also dedicated to the Xenopus laevis oocyte system.

Individual cells behave in surpnsmg ways that cannot be deduced from the averaged results of an organ as assessed by the use of conventional biochemical methods. Thus multicellular plant and animals systems are being investigated by an increasing array of histochemical and cytochemical techniques based on general chemical or specific immunological interactions to identify structural materials and to assess biological activities. In recent years there has been an increasing range of fluorescent probes, along with advanced computerised imaging and analysis techniques, which allows the behaviour of individual living cells to be followed in considerable detail. The parallel use of microinjection, microelectrodes and patch-clamping provides additional information about cells and their responses. Recombinant DNA technology has highlighted the desirability and the power of microinjecting defined materials into specific cells and so manipulating their fundamental biochemistry. New hypotheses are being tested which will form the cornerstone of future developments across the whole spectrum of biotechnology. The First European Workshop on Biotechnology Applications of Microinjection, Microscopic Imaging and Fluorescence was run at the University of East London, U.K, 21st-24th April, 1992 with the objective of bringing together a diverse group of individuals who were using these state-of-the-art applications for biotechnological exploration. A novel feature of the meeting was paiticipation by instrument manufacturers in the programme: there were hands-on workshops (where living cells could be examined), combined with the poster sessions.

Current Topics in Membranes and Transport

By combining the tools of organic chemistry with those of physical biochemistry and cell biology, Non-Natural Amino Acids aims to provide fundamental insights into how proteins work within the context of complex biological systems of biomedical interest. The critically acclaimed laboratory standard for 40 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. With more than 400 volumes published, each Methods in Enzymology volume presents material that is relevant in today's labs -- truly an essential publication for researchers in all fields of life sciences. - Demonstrates how the tools and principles of chemistry combined with the molecules and processes of living cells can be combined to create molecules with new properties and functions found neither in nature nor in the test tube - Presents new insights into the molecular mechanisms of complex biological and chemical systems that can be gained by studying the structure and function of non-natural molecules - Provides a "one-stop shop" for tried and tested essential techniques, eliminating the need to wade through untested or unreliable methods