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Microanatomical Labeling of Germinal Center Structures for Flow Cytometry Using Photoactivation

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Book cover Germinal Centers

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1623))

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.

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References

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Correspondence to Gabriel D. Victora .

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Jacobsen, J.T., Victora, G.D. (2017). Microanatomical Labeling of Germinal Center Structures for Flow Cytometry Using Photoactivation. In: Calado, D. (eds) Germinal Centers. Methods in Molecular Biology, vol 1623. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7095-7_4

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  • DOI: https://doi.org/10.1007/978-1-4939-7095-7_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7094-0

  • Online ISBN: 978-1-4939-7095-7

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