Counterstaining Protocols

DAPI Counterstaining

1. Wash and equilibrate the sections with PBS for 5 min*3.
2. Dilute the DAPI solution to an appropriate concentration of working solution as the manual instruction.
3. Drop appropriate amount of DAPI working solution to the slides as the manual instruction (Normally 300-400µl) and make sure the tissue is all covered by the solution.
4. Incubate at room temperature for 5 min.
5. Wash in PBS bath for 5 min*3.
6. Mount the sections with mounting medium and coverslip.
7. To make this step simple, mounting medium (DAPI-contained) is recommended.

Haematoxylin Eosin (H&E) Counterstaining

1. Counterstain in eosin-phloxine solution for 30 seconds to 1 minute.
2. Dehydrate through 95% alcohol, 2 changes of absolute alcohol, 5 minutes each.
3. Clear in 2 changes of xylene, 5 minutes each.
4. Mount with xylene based mounting medium.