Handbook of ELISPOT: Methods and Protocols / Edition 2

by Alexander E. Kalyuzhny

Overview

Enzyme-linked immunospot assay (ELISPOT) has been known for some time as a unique state-of-the-art technique for studying the cytokine-secreting activity of immune system cells, and it appears to be one of the fast growing applications in biomedical research, becoming an indispensable tool in vaccine development, HIV research, transplantation studies, and cancer and allergy research. The second edition of Handbook of ELISPOT: Methods and Prools, only the second book in the field which is entirely dedicated to ELISPOT assay, shares the detailed techniques that have been developed since the release of the popular first edition. Straight from the labs of seasoned experts, this book covers setting and performing ELISPOT assays, ELISPOT for veterinary research, advanced ELISPOT techniques, image and data analysis, as well as vaccine development and diagnostics. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective chapters, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory prools, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Handbook of ELISPOT: Methods and Prools, Second Edition serves as a compilation of a technical reference and a troubleshooting guide for researchers, both experienced and novice, worldwide in order to advance the usage of this key tool.

Product Details

ISBN-13:	9781493961115
Publisher:	Springer-Verlag New York, LLC
Publication date:	08/23/2016
Series:	Methods in Molecular Biology , #792
Edition description:	Softcover reprint of the original 2nd ed. 2012
Pages:	261
Product dimensions:	7.01(w) x 10.00(h) x (d)

Table of Contents

Unique Strengths of ELISPOT for T Cell Diagnostics.- The Impact of Harmonization on ELISPOT Assay Performance.- Equine ELISPOT Assay to Study Secretion of IFNγ and IL-4 from Peripheral Blood Mononuclear Cells.- Utilization of Feline ELISPOT for Mapping Vaccine Epitopes.- Analyzing Cellular Immunity to AAV in a Canine Model Using ELISPOT Assay.- Dual and Triple Color Fluorospot.- ELISPOT Assay as a Tool to Study Oxidative Stress in Lymphocytes.- ELISPOT Assay for Neuroscience Research: Studying TNFa Secretion from Microglial Cells.- ELISPOT Assay as a Tool to Study the Effects of Stem Cells on Cytokine Secretion.- Combining ELISPOT and ELISA to Measure Amounts of Cytokines Secreted by a Single Cell.- How ELISPOT Morphology Reflects on the Productivity and Kinetics of Cells’ Secretory Activity.- Mathematical Algorithms for Automatic Search, Recognition, and Detection of Spots in ELISPOT Assay.- Objective, User-Independent ELISPOT Data Analysis Based on Scientifically Validated Principles.- Statistical Analysis of ELISPOT Assays.- Response Determination Criteria for ELISPOT: Toward a Standard that Can Be Applied Across Laboratories.- Detection of Vaccinia Virus-Specific IFNγ and IL-10 Secretion from Human PBMCs and CD8⁺ T Cells by ELISPOT.- ELISPOT Assays to Enumerate Bovine IFN-γ Secreting Cells for the Development of Novel Vaccines Against Bovine Tuberculosis.- IL-7 Addition Increases Spot Size and Number as Measured by T-SPOT.TB®.- Overview of Membranes and Membrane Plates Used in Research and Diagnostic ELISPOT Assays.

Editorial Reviews

From the reviews of the second edition:

“In this second edition … the reader is introduced not only to the basic concept behind the technique but also to many different applications in common use. Importantly, the book serves as a reference guide with lots of trouble shooting and discussion of common problems. … This text would be key reading material for any laboratory planning to set up this technique. … this text is comprehensive and detailed with a good use of raw data and formula to make following the prools simple.” (Hayley Evans, Immunology News, November, 2012)

From the Publisher

Reviewer: Marion C. Cohen, PhD (SUNY Downstate Medical Center)
Description: This book is part of the Methods in Molecular Biology series that is designed to provide descriptions and applications of specific laboratory techniques to both neophytes as well as experienced researchers. This volume is devoted to the use of the ELISPOT assay, a technique used to quantify cytokine-secreting cells in many fields of biomedical research.
Purpose: According to the editor, this is the first book devoted entirely to the ELISPOT assay. Although it has been used for more than 20 years, the assay remains a challenge to set up and interpret the results. The book is expected to serve as a reference manual for beginners and as a troubleshooting guide for experienced ELISPOT users. It includes a collection of protocols covering a variety of topics which could be adapted by almost any potential user. In addition it begins with a history of the assay which is a nice touch.
Audience: The target audience includes anyone who may wish to use the ELISPOT assay. The assay has been used in many areas of research including vaccine development, transplantation studies, HIV, cancer, and allergy research as well as studies that focus on basic immunologic mechanisms. The quantification of cytokine-secreting cells is important for a large variety of areas of research. The editor is an authority in the field and the individual chapters have been written by experts.
Features: The book begins with the history behind the development of the assay. Chapters 2-8 cover how to set up the assay and evaluate the results. This is followed by application of ELISPOT assays to animal and human studies (chapters 9-16). Chapters 17-20 describe modifications of the assay. The techniques are well described with notes that offer helpful information. Most of the figures are black and white but 11 of them are also in color and are placed together in one section with identification of the chapter to which each belongs. There are appropriate references and an index.
Assessment: Although techniques are often better learned via a hands-on approach, this book would be a good way to develop the ELISPOT assay for one's own laboratory if there were no one available to demonstrate it. The protocols are detailed and the helpful hints in the form of "notes" are useful. The early chapters supply a wealth of detail on the setup and rationale behind aspects of the technique. Even if one has experience with the technique, there are modifications that might be helpful. And if all else fails, the authors of each chapter provide a potential contact source for problem solving.

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