Dfc280 plus camera

After creating the groups, the slides were fixed in 4% paraformaldehyde for about 30 min. Then the slides were washed twice in phosphate-buffered saline (PBS) solution for 15 min. The endogenous peroxidase activity was blocked with 3% hydrogen peroxide (Fisher Scientific, Melrose Park, IL, USA) (diluted with PBS) for 15 min and the slides were washed three times in PBS solution. The epitopes were stabilized by applying serum blocking solution for 15 min. The slides were incubated with the anti-leptin receptor primary antibody for 60 min at room temperature. The slides were washed three times in PBS solution, after which the biotinylated secondary antibody was applied for 10 min. The slides were then washed three times in PBS solution. Thereafter, streptavidin peroxidase was applied to the slides for 10 min. After washing with PBS, 3,3’-diaminobenzidine (DAB) was used as a chromogen. Then all the slides were counterstained with Harris’s hematoxylin. The slides were examined with a photo-light microscope (DM4000B Image Analysis System, Leica, Germany) and a Leica DFC280 Plus camera.