This volume provides key methods and protocols from laboratories engaged in germinal centers (GC) research with the expectation of stimulating further research, and to aid scientists in the study of GC biology and pathology. 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.Authoritative and practical, Germinal Centers: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Includes cutting-edge methods and protocols

Analysis of the Germinal Center Reaction in Tissue Sections.- Detecting Gene Expression in Lymphoid Microenvironments by Laser Microdissection and Quantitative RT-PCR.- Tracking Plasma Cell Differentiation in Living Mice with 2-Photon Microscopy.- Microanatomical Labeling of Germinal Center Structures for Flow Cytometry Using Photoactivation.- Fate Mapping and Transcript Profiling of Germinal Center Cells by Two-photon Photoconversion.- Intravital Microscopy of T-B Cell Interactions in Germinal Centers.- Identifying Follicular Regulatory T cells by Confocal Microscopy.- Cytokine Expression by T follicular Helper Cells.- Follicular Dendritic Cell Isolation and Loading of Immune Complexes.- Isolation and Characterization of Mouse and Human Follicular Dendritic Cells.- In-vitro-induced Germinal Center B Cell Culture System.- CRISPR/Cas9 Mediated In Vitro Mutagenesis in GC-like B Cells.- Germinal Center Formation with Retrovirally-transduced B Cells for Determining the Role of Specific Molecules In Vivo.- Characterization of the B Cell Transcriptome by RNA-Binding Proteins using iCLIP.- Flow-Cytometric Method Measuring B Cell Surface Immunoglobulin Avidity.- Somatic Hypermutation and Affinity Maturation Analysis Using the 4-hydroxy-3-nitrophenyl-acetyl (NP) System.- Targeting Gene Function in Germinal Center B Cells: A Practical Approach.- Development of Mouse Model Systems of Germinal Centre Lymphomas.- The AID-Cre-ERT2 Model: A Tool for Monitoring B Cell Immune Responses and Generating Selective Hybridomas.- Determining the Origin of Human Germinal Center B Cell-derived Malignancies.- Tracking B-cell Repertoires and Clonal Histories in Normal and Malignant Lymphocytes.- How to Simulate a Germinal Centre.