Immunofluorescence Imaging of Liver Macrophages

Liver tissues were cryo-sectioned and processed for the immunofluorescence (IF) assay as described previously (Alam et al., 2012 (link)). RBC-EM ^DiD or PBS-injected mouse liver sections were stained with anti-rabbit CD68 (Abcam), followed by staining with goat anti-rabbit FITC (Abcam). Tissue sections were mounted using the VECTASHIELD mounting medium (Vector Laboratories, Burlingame, CA, United States). IF-stained sections were imaged under a confocal microscope (LSM 5 Exciter, Zeiss, Oberkochen, Germany). Total of six fields were counted by observers. Number of CD68 positive (CD68 +; green) cells were counted from RBC-EM ^DiD or PBS-injected mouse liver sections. Then the number of DiD positive (DiD +; red) with CD68 + or CD68 - cells were counted.