| Detachment of cover glasses |
| 1. The cover glass is removed by immersion in xylene (heating at 37°C or 60°C overnight when necessary). |
| 2. The mounting agent is removed using new xylene. |
1 hr×3 times |
| 3. 100% ethanol (removal of xylene) |
15 min×2 times |
| 4. Specimens are passed through a descending ethanol series (95-70%) for 5 minutes each, and washed with water (2 min). |
| Decolorization of old Pap specimens |
| Specimens are treated with an optical clearing agent (SCALEVIEW-A2) for 5-10 minutes and placed in phosphate buffer (pH, 7.2) to terminate the reaction of the clearing agent. |
| Pap staining |
| Basically, the routine staining method is used. However, it is recommended to use a longer time (30-40 dips) for differentiation after hematoxylin staining. |
