HistoLogic Stainer is designed to run IHC, IHC-P, IF and mIF.

Quick Protocol for IHC-P (for detailed instructions, see the PDF Manual)

A: Prepare the Slides

1. Cut the sections and heat them at 60C for 60 min.

2. Let cool down and keep them in the fridge until ready for IHC-P.

3. Take the slides out of fridge, label them from 1-32 with Pencil.

4. Melt the wax at 60C for 5 min.

5. Dewax and rehydrate the sections and place the slides in Tap water until ready for HIER.

6. Do the HIER using Tris-EDTA, pH 9.00.

B: IHC-P on HistoLogic Stainer - Simultaneous staining of 32 slides in less than 60 min.

1. Load the HistoLift with 32 slides: slide 1 is placed in Position 1 etc.

2. Place the HistoLift in HistoRinse, dip 3x.

3. Place the HistoLift in Peroxide Block for 5 min.

4. Place the HistoLift in HistoRinse, dip 3x.

5. Place the HistoLift in Protein Block for 5 min.

6. Place the HistoLift in HistoWash, dip 3x.

7. Place the HistoLift in HistoCell-Primary Ab for 15 min.

8. Place the HistoLift in HistoRinse, dip 3x 5s with a gentle agitation.

9. Place the HistoLift in HistoWash, dip 3x 5s with a gentle agitation.

10. {Place the HistoLift in HistoCell-AdaptoBody for 15 min (if required).}

11. Place the HistoLift in HistoRinse, dip 3x 5s with a gentle agitation.

12. Place the HistoLift in HistoWash, dip 3x 5s with a gentle agitation.

13. Place the HistoLift in HistoCell-Secondary Ab for 15 min (Polymer can be diluted1:3).

14. Place the HistoLift in HistoRinse, dip 3x 5s with a gentle agitation.

15. Place the HistoLift in HistoWash, dip 3x 5s with a gentle agitation.

C: Stain

1. Take the slides out of HistoLift and place them on Osenses’ HistoTray.

2. Add the Osenses’ Candela DAB chromogen and wait for 5 min.

3. Counterstain with Harris Hematoxylin.

4. Coverslip and let dry on HistoTray.

D: Clean-up & reuse the reagents

Wash the HistoCells and HistoVoirs with tap water, followed by a final rinse with deionized (DI) water.

Store all reagents in the fridge either inside the HistoCells or in another suitable container for future use.