Skin sections were stained with H&E and the epidermal thickness was determined by measuring the average interfollicular distance under the microscope in a blinded manner. For IHC staining, skin cryosections were fixed, blocked, and stained with rat-anti-mouse Gr-1 mAb (1:50) followed by goat-anti-rat IgG secondary Ab (1:200, Southern Biotech) 4 (link). Slides were developed with 3-amino-9-ethylcarbazole (AEC) substrate solution (Vector Laboratories) and counterstained with hematoxylin. Images were acquired at x200 magnification using Aperio ScanScope digital scanners.
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Epidermal Thickness and Immune Cell Analysis
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Corresponding Organization :
Other organizations : University of Louisville, Ruijin Hospital, Pennsylvania State University
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Variable analysis
independent variables
- Staining method (H&E staining, IHC staining with rat-anti-mouse Gr-1 mAb)
- Epidermal thickness (measured as average interfollicular distance)
- Gr-1 positive cell staining (visualized with AEC substrate solution)
- Blinding (epidermal thickness measurements)
- Microscope magnification (x200)
- Positive control: None specified
- Negative control: None specified
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