This fifth edition volume expands on the previous editions by presenting readers with the latest developments and emerging methodologies in cytometry. The chapters in this book cover cytometry basics such as lasers for cytometry, metrics that can be used to evaluate spillover spreading, and the process of panel design and iterative optimization for spectral flow cytometry; novel methodologies such as image-enabled cell sorting, co-staining of fluorochrome-conjugated and oligonucleotide-conjugated antibodies, and screening for cell type selective probes; and a look at the achievements made in the clinical setting for both flow and mass cytometry. 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, readily reproducible step-by-step laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and comprehensive, Flow Cytometry Protocols, Fifth Edition is a valuable resource for researchers and scientists who are interested in continuing or expanding their knowledge of this developing field.