Hoechst Blue Stain

Staining of FFPE tissue section

• Deparaffinize FFPE sections with Xylene.
• Rehydrate sections with decreasing concentrations of ethyl alcohol (100% to 70%).
• Wash slides 2 times with dH₂O.
• Dip slides into universal antigen retrieval buffer.
• Microwave sections at boiling temperature for 10min.
• Wash slides 2 times with dH₂O.
• Dip slides into FFPE autofluorescence quenching buffer for 20min.
• Wash slides with 70% ethyl alcohol.
• Wash slides with 1xPBS.
• Drain Wash solution against a towel and put slides into the wet chamber.
• Dilute Hoechst Blue Stain at 1:100 in Block solution and add to each slide (300-500 µl is sufficient to cover the section on each slide).
• Incubate the slides in the wet chamber at room temperature for 1hr.
• Take slides out of the wet chamber, drain off Hoechst Blue Stain, and wash three times with the Wash solution.
• Add 20 ul of Mowiol solution to the section on each slide. Place a cover glass onto the section and apply a small pressure to remove any trapped air bubble. Let Mowiol solution dry at room temperature for 1hr. Store glass slides at 4^oC for less than 1 month.