Kling on hier

Tissues fixed in 10% buffered formalin were processed for analysis by the conventional technique for histology and stained with hematoxylin and eosin. For immunohistochemistry, samples were processed using the streptavidin–biotin–peroxidase complex (Histostain-Plus, Invitrogen, USA) as previously described, with slight modifications (Valenzuela-Moreno et al., 2022a ). Briefly, paraffin-embedded sections were cut and mounted on electrocharged slides (Kling-On-Hier, Biocare). Subsequently, the tissues were incubated with serum from T. gondii-positive mice experimentally infected with the T. gondii Me49 strain (dilution 1:300). The slides were then incubated with a secondary multispecies biotinylated antibody (Invitrogen, USA), followed by incubation with streptavidin-peroxidase. Immune complexes were revealed with the commercial Betazoid DAB solution, chromogen 3,3′ diaminobenzidine (Biocare®). Liver and spleen sections of mice infected with T. gondii strain Me49 were used as positive controls, and the primary antibody was replaced with PBS as a negative control. Histological and IHC sections were examined by light microscopy (Zeiss Axiostar plus; Göttingen, Germany).

Representative samples of fixed tissues were processed following the routine histological protocol and stained with hematoxylin and eosin. Immunoreactivity was visualized by the streptavidin-biotin-peroxidase complex (Histostain-Plus, Invitrogen, USA), as previously described with slight modifications (11 ). Briefly, paraffin-embedded sections were cut and mounted in electrocharged slides (Kling- On- Hier, Biocare); the tissues were deparaffined, hydrated, and blocked following the routine methodology. After that, slides were incubated T. gondii positive serum by ELISA, from a mouse experimentally infected with T. gondii Me49 strain (1:400 dilution). All slides were washed and incubated with a secondary multispecies biotinylated antibody (Invitrogen, USA), followed by an incubation step with streptavidin-peroxidase. Immunocomplexes were revealed with a commercial solution (Betazoid DAB, Chromogen 3, 3' Diaminobenzidine, Biocare^®). Sections of liver and spleen of infected mice with T. gondii Me49 strain were used as positive controls and for the negative control the primary antibody was replaced with PBS. Histopathology and IHC slides were examined by optical microscopy (Zeiss Axiostar plus; Göttingen, Germany).