Immunohistochemestry vs. Immunofluorescence: Comparative analysis via software of total colorimetric reaction of GPR43 protein in adipose tissue

Immunohistochemistry revealed by diaminobenzidine (IHC-DAB) or fluorescence (IHC-F) are two of the most common techniques used in histopathology. In order to evaluate the similarity between two methodologies that share the same antigen-antibody reaction principle, using Image-Pro Plus software we performed total colorimetry quantification and comparison of immunohistochemical reaction; revealed with diaminobenzidine (IHC-DAB) and FITC fluorescence (IHC-F). As biological model, GPR43 protein was assessed by Immunohistochemestry in inguinal, mesenteric and gonadal (WAT) cd1 mice adipose tissue. In predefined areas of digital images, we evaluated the reaction obtained following the same processing conditions. Results: The GPR43 protein in both IHC-DAB and IHC-F was evidenced in adipocytes membrane and infiltrated immune cells. The average of positive reaction in μm² (total colorimetry x μm²) on a predefined area of 25x10⁴ μm², showed no significant differences in the distinct tissues between the performed methods. Conclusions: In the total colorimetric analysis between the two methodologies IHC-DAB and IHC-F, the resulting quantitative values were similar for all adipose tissue compartments studied.