Digital camera dsc 85

Piscidin1 was examined with an optical microscope and immunohistochemical techniques. Slices were treated with an anti-Piscidin1 antibody overnight in a humid environment. Following a PBS wash, the sections were incubated with a goat anti-rabbit IgG-peroxidase conjugate for 60 min (Sigma-Aldrich, St. Louis, MO, USA, dilution 1:100, source Goat). By allowing the sections to sit in a solution of 0.02% diaminobenzidine (DAB) and 0.015% hydrogen peroxide for 1–5 min at room temperature, the peroxidase activity of the sections was assessed. Sections were dehydrated, mounted, and inspected with a Zeiss Axioskop 2 plus microscope and a Sony Digital Camera DSC-85 after being rinsed in PBS. As a negative control, experiments were conducted without the primary antibody.

Immunohistochemical procedures and an optical microscope were used to evaluate S100 (Sigma-Aldrich, St. Louis, MO, USA, dilution 1:100, source Rabbit). In a humidified atmosphere, slices were treated overnight with S100 antibody. The sections were then washed in PBS and incubated for 60 min with a goat anti-rabbit IgG-peroxidase conjugate (Sigma-Aldrich, St. Louis, MO, USA, dilution 1:100, source Goat) from Sigma-Aldrich. The peroxidase activity of the sections was determined by incubating them in a solution of 0.02% diaminobenzidine (DAB) and 0.015% hydrogen peroxide for 1–5 min at room temperature. [58 (link)]. After being rinsed in PBS, sections were dehydrated, mounted, and viewed with a Zeiss Axioskop 2 plus microscope and a Sony Digital Camera DSC-85. Experiments were carried out without the main antibody as a negative control.

Immunohistochemical techniques, testing TLR‐2, VIP with a light microscope for observation. Sections were incubated overnight in a humid chamber with the following antibodies: TLR2 (Toll‐like Receptor 2 Antibody, product in rabbit by Active Motif, La Hulpe, Belgium, Europe, 1:125) and VIP (Vasoactive intestinal polypeptide, product in rabbit by Sigma‐Aldrich, St. Louis, MO, 1:4000). Then, the sections were washed in phosphate‐buffered saline (PBS) and incubated for 60 min with a goat anti‐rabbit IgG‐peroxidase conjugate. Peroxidase activity was determined by incubating the sections in a solution of 0.02% diaminobenzidine (DAB) and 0.015% hydrogen peroxide for 1–5 min at room temperature (Lauriano et al., 2015 (link); Zaccone, Icardo, et al., 2017 (link)). After rinsing in PBS, sections were dehydrated, mounted, and examined under a Zeiss Axioskop 2 plus microscope equipped with a Sony Digital Camera DSC‐85. Control experiments excluding primary antibody were performed (data not showed).