Formalin-fixed and paraffin-embedded specimens were sectioned at 5 μm thickness and mounted on glass slides. The PD-L1 (clone 5H1) antibody was obtained from Lieping Chen (12 (link)). In brief, antigen retrieval was achieved with citrate buffer. Anti-PD-L1/B7-H1 murine IgG (1:1000) was applied on tissue sections overnight at 4°C. After washing with TBS (0.05 M Tris base, 0.9% NaCl, pH 8.4), tissue sections were incubated with biotinylated anti-mouse IgG (1:100; BA-2001, Vector Laboratories) followed by incubation with Elite ABC kit for 30 min at room temperature. Antibody binding was detected with the TSA Biotin System (NEL700A001KT, PerkinElmer) and DAB (DAKO, K3468). Murine PD-L1 staining was performed using the polyclonal antibody against mouse B7-H1/PD-L1 antibody (1:100, R&D Systems). Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and KIT immunostaining (1:200, D13A2; Cell Signaling Technology) were performed as described (24 (link)). Slides were analyzed and imaged on the Axio Imager 2 wide-field microscope (Zeiss).
Axio imager 2 wide field microscope
Manufactured by Zeiss
The Axio Imager 2 is a wide-field microscope designed and manufactured by Zeiss. It is equipped with advanced optical components and illumination systems to provide high-quality imaging and analysis capabilities. The core function of this microscope is to enable detailed observation and examination of various samples across multiple research and industrial applications.
Automatically generated - may contain errors
Lab products found in correlation
Biotinylated anti mouse igg, by Vector Laboratories (1 mentions)
Abc elite kit, by Vector Laboratories (1 mentions)
3 3 diaminobenzidine (dab), by Agilent Technologies (1 mentions)
Tsa biotin system, by PerkinElmer (1 mentions)
Clone bm8, by BioLegend (1 mentions)
Clone afs98, by BioLegend (1 mentions)
2 protocols using axio imager 2 wide field microscope
1
Immunohistochemical Analysis of PD-L1 Expression
2
Immunohistochemical Analysis of Liver Macrophages
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Liver specimens were fixed in 4% paraformaldehyde, embedded in paraffin, and sectioned to 5 μm thickness. Antigen retrieval was achieved with citrate buffer. Tissue sections were stained overnight at 4°C with anti–mouse F4/80 (1:100 dilution; clone BM8), Csf1r (1:200 dilution; clone AFS98; both BioLegend), or Ki-67 (1:200 dilution; clone 16A8; Vector Laboratories). After being washed with PBS, slides were incubated with a fluorochrome-conjugated secondary antibody and then analyzed and imaged on an Axio Imager 2 wide-field microscope (Zeiss).
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