In this first book dedicated entirely to the ELISPOT, a critical enzyme-linked immunospot assay used widely in biomedical research, recognized experts with first-hand experience detail how to design, perform, and analyze these assays. The readily reproducible techniques they provide cover a wide variety of topics, including the use of membrane-backed plates, the standardization and validation procedures, the removal of cells from ELISPOT plates, cell separation techniques, and the quantification of ELISPOT data. There are also numerous ELISPOT applications involving animal models, human cells, measles, multiple sclerosis, immune responses, multicytokine detection systems, and immunocytochemistry. Highlights include dual-color and multiplex ELISPOT assays, use of the ELISPOT assay on feline lymphocytes, standardization of the ELISPOT procedure, and combining the ELISPOT assay with immunohistochemistry.

In this first book dedicated entirely to the ELISPOT, a critical enzyme-linked immunospot assay used widely in biomedical research, recognized experts with first-hand experience detail how to design, perform, and analyze these assays. The readily reproducible techniques they provide cover a wide variety of topics, including the use of membrane-backed plates, the standardization and validation procedures, the removal of cells from ELISPOT plates, cell separation techniques, and the quantification of ELISPOT data. There are also numerous ELISPOT applications involving animal models, human cells, measles, multiple sclerosis, immune responses, multicytokine detection systems, and immunocytochemistry. Highlights include dual-color and multiplex ELISPOT assays, use of the ELISPOT assay on feline lymphocytes, standardization of the ELISPOT procedure, and combining the ELISPOT assay with immunohistochemistry.

This first edition volume demystifies the complex topic of flow cytometry by providing detailed explanations and nearly 120 figures to help novice flow cytometry users learn and understand the bedrock principles necessary to perform basic flow cytometry experiments correctly. The book divides the topic of flow cytometry into easy to understand sections and covers topics such as the physics behind flow cytometry, flow cytometry lingo, designing flow cytometry experiments and choosing appropriate fluorochromes, compensation, sample preparation and controls and ways to assess cellular function using a variety of flow cytometry assays. Written as a series of chapters whose concepts sequentially build off one another, using the list of materials contained within each section along with the readily reproducible laboratory protocols and tips on troubleshooting that are included, readers should be able to reproduce the data figures presented throughout the book on their way to mastering sound basic flow cytometry techniques. Easy to understand and comprehensive, Flow Cytometry Basics for the Non-Expert will be a valuable resource to novice flow cytometry users as well as experts in other biomedical research fields who need to familiarize themselves with a basic understanding of how to perform flow cytometry and interpret flow cytometry data. This book is written for both scientists and non-scientists in academia, government, biotechnology, and medicine.

This book provides the immune oncology (IO) community with a deeper understanding of the scope of the biomarker methods to potentially improve the outcome from immunotherapy. The editors secured the input from experts in the field dedicated to translating scientific research from bench to bedside was submitted. The book provides not only details about the technical, standardization and interpretation aspects of the methods but also introduces the reader to the background information and scientific justification for selected biomarkers and assays. Written in the highly 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Since the publication of the first edition of the Handbook of Human Immunology in 1997, major scientific achievements have directly contributed to an increased understanding of the complexities of the human immune system in health and disease. Whether as a result of the sequencing of the entire human genome, or of technological advancements, severa

The recent explosion of information in innate immune pathways for recognition, effect or responses, and genetic regulation has given impetus to investigations into analogous pathways in the human immune response, which in turn has produced attendant insights into both normal physiology and immunopathology. This volume presents a compendium of methods and protocols for the investigation of human innate immunity with application to the study of normal immune function, immunosenescence, autoimmunity and infectious diseases. Among the topics covered are quantitative flow cytometry for Toll-like receptor expression and function; multidimensional single cell mass cytometry (CyTOF) in complex immune interactions and tumor immunity; imaging techniques such as Imagestream high resolution microscopy coupled to flow cytometry, immune cell infiltration of organotypic, biomimetic organs; high-throughput single cell secretion profiling; multiplexed transcriptomic profiling; microsatellite and microRNA methodologies, RNA interference; and the latest bioinformatics and biostatistical methodologies, including in-depth statistical modeling, genetic mapping, and systems approaches.