After PET imaging, animals were sacrificed and tissue in the region of LPS/PBS injections were dissected, placed in 4% paraformaldehyde for overnight incubation, and transferred to 30% sucrose solution overnight. Subsequently they were embedded in O.C.T. medium (Fischer Scientific, Waltham, MA, USA) and frozen overnight at −80 degrees. Samples were sectioned on a cryostat at 5–10-micron sections.
Anti-Granzyme B (Cat #4059, Abcam, Cambridge, MA, USA) immunofluorescence was performed according to a standard protocol [21 (link)] using Anti-Granzyme B primary antibody 1:100 and a secondary goat anti-rabbit antibody 1:1000 (Thermo Fisher, Waltham, MA, USA). Slides were imaged on an Echo Revolve 332 microscope (Echo) at 4×, 10×, and 40× magnification.
Anti-Granzyme B (Cat #4059, Abcam, Cambridge, MA, USA) immunofluorescence was performed according to a standard protocol [21 (link)] using Anti-Granzyme B primary antibody 1:100 and a secondary goat anti-rabbit antibody 1:1000 (Thermo Fisher, Waltham, MA, USA). Slides were imaged on an Echo Revolve 332 microscope (Echo) at 4×, 10×, and 40× magnification.
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