Mouse liver tissues were fixed in 10% buffered formalin for 24 h at room temperature. Formalin-fixed, paraffin-embedded sections were cut with thickness of 5-μm. H&E staining was performed on deparaffinized and rehydrated sections according to manufacturer’s protocol (Leica Biosystems, Buffalo Grove, IL, USA). Immunochemistry for F4/80 and HA staining were carried out by established procedures as previously described (Yang et al. 2019 (link)). F4/80 monoclonal antibody or biotin-labeled HA-binding protein were applied to mouse liver sections. VECTASTAIN Elite ABC kit and DAB Peroxidase Substrate kits were used as directed by the manufacturer (Vector Laboratories, Burlingame, CA, USA). HA-positive area was evaluated from randomly selected 10 fields of × 200 magnification per slide and quantified with NIH Image J software.
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Histological Analysis of Mouse Liver
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Biotin binding protein
Buffalo
Formalin
Liver
Monoclonal antibody
Mouse
Paraffin
Peroxidase
Tissues
Vector
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Variable analysis
independent variables
- Fixation of mouse liver tissues in 10% buffered formalin for 24 h at room temperature
- Staining techniques: H&E staining, F4/80 immunochemistry, and HA staining
- F4/80-positive area
- HA-positive area
- Thickness of formalin-fixed, paraffin-embedded sections (5-μm)
- Deparaffinization and rehydration of sections
- Use of established procedures for F4/80 and HA staining as previously described (Yang et al. 2019)
- Use of VECTASTAIN Elite ABC kit and DAB Peroxidase Substrate kits as per manufacturer's instructions
- Evaluation of HA-positive area from randomly selected 10 fields of × 200 magnification per slide
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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