Histology and Immunohistochemistry

1. The skin tissues were fixed in 4% paraformaldehyde fixation solution.
2. Embed skin tissues with paraffin blocks for routine sectioning.
3. The sections were successively dewaxed and inactivated with 3% H2O2.
4. Blocked with 5% BSA at room temperature for 30 min.
5. Washed with PBS for three times.
6. Remove PBS and add primary antibodies in block solution for 24 hours at 4 ⁰C on a shaker.
7. Remove primary antibodies and wash sections with PBS for three times.
8. Remove PBS and add species-specific Alexa fluorophore-conjugated secondary antibodies in block solution at room temperature for 30 mins. 
9. Remove secondary antibodies and wash sections in PBS for three times.
10. The sections were observed and photographed using a microscope connected to a digital camera.
 

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