Fate Mapping and Transcript Profiling of Germinal Center Cells by Two-Photon Photoconversion
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 âsnapshotâ 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 âtaggingâ 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.
|Authors||Moran, I.; Phan, T.|
|Responsible Garvan Author|
|Publisher Name||Methods Mol Biol|
|URL link to publisher's version||https://www.ncbi.nlm.nih.gov/pubmed/28589347|