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Microanatomical Labeling of Germinal Center Structures for Flow Cytometry Using Photoactivation.
- Source :
-
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2017; Vol. 1623, pp. 51-58. - Publication Year :
- 2017
-
Abstract
- Germinal centers (GCs) are inducible microanatomical structures required for the generation of high-affinity antibodies. Migration of B and T cells within and into/out of GCs plays a key role in the evolutionary process that underlies affinity maturation, and thus microanatomical location of cells is an important variable when studying GC processes. We describe a protocol in which in situ photoactivation by multiphoton microscopy can be used to add microanatomical information to flow cytometry, allowing for identification and isolation of GC cells based on their location. Cells in different microanatomical compartments can then be sorted and analyzed for surface marker and mRNA expression.
- Subjects :
- Animals
B-Lymphocytes cytology
B-Lymphocytes immunology
B-Lymphocytes metabolism
Biomarkers
Germinal Center immunology
Lymph Nodes immunology
Lymph Nodes metabolism
Mice
Microscopy, Fluorescence, Multiphoton
Cellular Microenvironment
Flow Cytometry methods
Germinal Center cytology
Germinal Center metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1940-6029
- Volume :
- 1623
- Database :
- MEDLINE
- Journal :
- Methods in molecular biology (Clifton, N.J.)
- Publication Type :
- Academic Journal
- Accession number :
- 28589346
- Full Text :
- https://doi.org/10.1007/978-1-4939-7095-7_4