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Fate Mapping and Transcript Profiling of Germinal Center Cells by Two-Photon Photoconversion

Abstract

The germinal center (GC) reaction is the key process for the generation of high affinity antibodies to foreign antigen. Standard experimental techniques such as fluorescence-activated cell sorting and histology have provided numerous insights into the composition and function of the GC. However, these approaches are limited to a “snapshot” in time and are unable to fully capture the dynamic nature of the GC. Intravital twophoton microscopy overcomes these disadvantages and has led to several major advances in the field but is restricted by practical and technical limits that prevent long-range mapping and molecular studies. Here we describe procedures for optical marking or “tagging” of cells in precise microanatomical compartments by two-photon photoconversion that can be used for long-term fate mapping and transcript profiling of GC T and B cells.

Type Journal
Authors Moran, I.; Phan, T.
Responsible Garvan Author A/Prof Tri Phan
Publisher Name Methods Mol Biol
Published Date 2017-06-07
Published Volume 1623
Published Pages 59-72
Status Published in-print
DOI 10.1007/978-1-4939-7095-7_5
URL link to publisher's version https://www.ncbi.nlm.nih.gov/pubmed/28589347