Immunofluorescence analysis of thymi was described previously 51 (link). Thymi were washed, fixed with 4% paraformaldehyde (PFA) for 1 h and snap frozen. 5-μm sections were blocked with PBS containing 5% bovine serum albumin (BSA) and goat serum (Jackson Laboratory) before staining. The sections were then covered with Prolong anti-fade mounting medium (Life Technologies) and images were obtained 1 to 3 d later with a Leica DM6000B Epi-Fluorescent microscope. Histo-cytometry analysis was performed as described previously 21 (link).
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Owen D.L., Mahmud S.A., Sjaastad L.E., Williams J.B., Spanier J.A., Simeonov D.R., Ruscher R., Huang W., Proekt I., Miller C.N., Hekim C., Jeschke J.C., Aggarwal P., Broeckel U., LaRue R.S., Henzler C.M., Alegre M.L., Anderson M.S., August A., Marson A., Zheng Y., Williams C.B, & Farrar M.A. (2019). Thymic regulatory T cells arise via two distinct developmental programs. Nature immunology, 20(2), 195-205.
Other organizations :
University of Minnesota Medical Center, Medical College of Wisconsin, University of Minnesota, University of California, San Francisco, Cornell University, University of Chicago, Salk Institute for Biological Studies
Thymi were fixed with 4% paraformaldehyde (PFA) for 1 h
Thymi were snap frozen
5-μm sections were blocked with PBS containing 5% bovine serum albumin (BSA) and goat serum (Jackson Laboratory) before staining
The sections were then covered with Prolong anti-fade mounting medium (Life Technologies)
positive controls
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negative controls
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