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 two-photon 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.
CITATION STYLE
Moran, I., & Phan, T. G. (2017). Fate mapping and transcript profiling of germinal center cells by two-photon photoconversion. In Methods in Molecular Biology (Vol. 1623, pp. 59–72). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7095-7_5
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