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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.
List of contents
Flow Cytometry: The Glass Is Half Full.- Basic Principles of Flow Cytometer Operation: The Make Your Own Flow Cytometer.- Lasers Sources for Traditional and Spectral Flow Cytometry.- How to Measure "Spillover Spread".- Comprehensive Immunophenotyping by Polychromatic Cytometry.- Panel Design and Optimization for Full Spectrum Flow Cytometry.- Practicalities of Cell Sorting.- Image-Enabled Cell Sorting Using the BD CellView Technology.- Monitoring Cell Proliferation by Dye Dilution: Considerations for Panel Design.- Multiparametric Analysis of Apoptosis by Flow Cytometry.- Quantitative and Standardized Pseudovirus Neutralization Assay for COVID-19.- CRISPR-Cas9-Mediated Bioluminescent Tagging of Endogenous Proteins by Fluorescent Protein-Assisted Cell Sorting.- Co-Staining with Fluorescent Antibodies and Antibody-Derived Tags for Cell Sorting Prior to CITE-seq.- Discovery of Live Cell Selective Fluorescent Probesand Elucidation of Their Mechanisms: Case Study of B Cell Selective Probe CDgB.- Fluorescence Lifetime Measurements and Analyses: Protocols Using Flow Cytometry and High-Throughput Microscopy.- Using Artificial Intelligence to Interpret Clinical Flow Cytometry Data Sets for Automated Disease Diagnosis and/or Monitoring.- Approaching Mass Cytometry Translational Studies by Experimental and Data Curation Settings.- CyTOF Intracellular Cytokine Assays for Antigen-Specific T Cells.- Imaging Mass Cytometry for In Situ Immune Profiling.- Cytometry in High-Containment Laboratories.
