1. Formalin fixed, paraffin-embedded tissue sections (3um thick) on silane-coated glass slides. (FRONTIER MATSUNAMI GLASS IND.,LTD OSAKA Japan)
2. Deparaffinize slides
3. The kitchen electric pot is filled with the Immunosaver diluted with distilled water by 200 times. (Fig.1)
4. The kitchen electric jar (pot) was boiled. (Fig.2)
5. Transfer the slides in staining racks into the pot and set the pot at keep warm temperature mode(98C) for 45 min. (Figs. 3.4)
6. Transfer the slides to PBS buffer solution. (Figs. 5.6)
(cool down is unnecessary.)
7. Rinse slides with PBS and start the Immunostaining protocol.