As previously described, a section of liver tissue was fixed in 4% formalin in PBS and embedded in paraffin using an avidin–biotin-peroxidase complex technique and fluorescent immunostaining after microwave antigen retrieval34 (link).
The sections were incubated at room temperature overnight with an anti-F4/80 antibody (1:250, Santa Cruz Biotechnology, Dallas, TX, USA) and successively treated with a biotinylated secondary antibody and peroxidase-avidin complex (ABC-kit; Vector Laboratories, Burlingame, CA, USA). Five images of the specimen were randomly separated and digitally captured at 400× magnification. The number of positive cells was then counted and assessed as the number per high-power field35 (link),36 (link).
The sections were incubated at room temperature overnight with an anti-F4/80 antibody (1:250, Santa Cruz Biotechnology, Dallas, TX, USA) and successively treated with a biotinylated secondary antibody and peroxidase-avidin complex (ABC-kit; Vector Laboratories, Burlingame, CA, USA). Five images of the specimen were randomly separated and digitally captured at 400× magnification. The number of positive cells was then counted and assessed as the number per high-power field35 (link),36 (link).
Free full text: Click here
