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.
Axioskop 2 plus microscope
Manufactured by Sony
Sourced in Germany
The Axioskop 2 plus microscope is a high-performance laboratory instrument designed for a wide range of microscopy applications. It features advanced optical components and a versatile design to enable precise and detailed observation of samples.
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4 protocols using axioskop 2 plus microscope
1
Immunohistochemical Analysis of Piscidin1
2
Immunohistochemical Analysis of Cell Signaling
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Analyses of MAPK p38, Piscidin1, and TLR2 were performed using an optical microscope and immunohistochemical techniques. Slices were exposed to anti-MAPK p38, anti-Piscidin1, and anti-TLR2 antibodies overnight in a humid environment. Slices were first washed in PBS and then incubated with a secondary antibody for 60 min. Slides were treated with diaminobenzidine (DAB) 0.02% and hydrogen peroxide 0.015% for a few minutes away from direct light. Sections were dehydrated, mounted, and evaluated using a Zeiss Axioskop 2 plus microscope (Oberkochen, Germany, Europe) and a Sony Digital Camera DSC-85 (Sony, Tokyo, Japan). As a negative control, experiments were conducted without the primary antibody.
3
Immunohistochemical Analysis of S100 Protein
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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.
4
Immunohistochemical Localization of TLR-2 and VIP
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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).
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