Flow Cytometry Basics for the Non-Expert (Techniques in Life Science and Biomedicine for the Non-Expert)

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. Front Matter ....Pages i-xvii Flow Cytometry: Definition, History, and Uses in Biological Research (Jody Bonnevier, Christopher Hammerbeck, Christine Goetz)....Pages 1-11 Physics of a Flow Cytometer (Jody Bonnevier, Jae-Bong Huh, Christopher Hammerbeck, Christine Goetz)....Pages 13-24 The Language of Flow Cytometry and Experimental Setup (Christine Goetz, Christopher Hammerbeck)....Pages 25-40 Fluorochrome Choices for Flow Cytometry (Christine Goetz, Christopher Hammerbeck)....Pages 41-51 Using the Process of Compensation to Prevent False Positive Data Caused by Fluorescence Spillover: A Practical Example (Christopher Hammerbeck, Christine Goetz, Li Jen Peng)....Pages 53-74 Primary and Secondary Antibodies and Flow Cytometry Controls (Christopher Hammerbeck, Christine Goetz, Jody Bonnevier)....Pages 75-102 Experimental Considerations with Data Sets as Examples (Christopher Hammerbeck, Christine Goetz, Li Jen Peng, Jae-Bong Huh)....Pages 103-148 Cell Enrichment (Christine Goetz, Christopher Hammerbeck, Andrew Wyman, Jae-Bong Huh)....Pages 149-155 Surface and Intracellular Staining Protocols for Flow Cytometry (Christine Goetz, Christopher Hammerbeck)....Pages 157-181 Troubleshooting (Christopher Hammerbeck, Christine Goetz)....Pages 183-203 Back Matter ....Pages 205-219 Flow cytometry allows for the measurement of different aspects of a cell or particles in real time. Applications include quantifying various types of T cells to determine the presence of various immune disorders, such as whether HIV has progressed to AIDS, studying the dynamics of immune signaling in cells to detect the suppression of normal immune function, and analyzing biopsy specimens for tumors, to aid in cancer diagnosis and prognosis. In addition to simplifying the technique and using vocabulary accessible to those not trained in cell biology and immunology, the book includes the following 4 unqiue features: 1) Tips on how to set up an experiment for flow cytometry optimially 2) An expanded data set of flow staining including multiparameter flow cytometry 3) Detailed staining protocols for flow cytometry (ex. protocols optimized for transcription factors, secreted cytokines, phospho-antibodies, and surface antigens) 4) Section devoted to antibody development and conjugation