1. Heat the sections at 60 °C for 60 minutes.
2. Immerse the sections into Xylene 5 min × 2times.
3. Immerse the sections into 100% EtOH 5min × 2times.
4. Immerse the sections into 85% EtOH 2 min.
5. Immerse the sections into 70% EtOH 2min.
6. Immerse the sections into 50% EtOH 2min..
7. Immerse the sections into 30% EtOH 2min.
8. Immerse the sections into 1X PBS 2min.
9. Incubate the sections with Antigen Retrieval Buffer at room temperature for 5 minutes.
10. Wash the sections with PBS Buffer 5 minutes × 3 times in a washing box.
11. Immerse the sections with Permeabilization Buffer at room temperature for 10 minutes.
12. Wash the sections with PBS Buffer 5 minutes × 3 times in a washing box.
13. Incubate the sections with Blocking Buffer at room temperature for 1 hour.
14. Incubate the sections with different antibodies in Primary Antibody Dilution Buffer at 4 °C overnight.
15. Wash the sections with PBS Buffer 5 minutes × 3 times in a washing box.
16. Incubate the sections with Fluor-conjugated secondary antibodies in Secondary Antibody Dilution Buffer at room temperature for 2 hours.
17. Wash the sections with PBS Buffer 5 minutes × 3 times in a washing box.
18. Rinse sections to be mounted with distilled water.
19. Tap the sections and then touch the edges of section on a paper to remove water.
20. Drop the top of sections with Mounting & Antifade Buffer with DAPI (Do Not Touch Tissue).
21. Detect with confocal microscope Or Store the slice in the dark at 4 °C.
