Mohs Mouse/Rabbit PolyDetector Plus DAB HRP Brown

Presentation

Specimen Preparation of Mohs Frozen Tissues

1. Embed the specimen in OCT inside a cryostat.
2. Cut sections at 4-5 µm and mount on a positively charged glass slide such as the Bio SB Hydrophilic Plus Slides (BSB 7028).
3. Air dry the slide at room temperature for 2 minutes and then incubate the slide at 60 °C for 3 minutes in an incubator or dry bath.
4. Fix in 100% acetone or 10% NBF for 2 minutes at room temperature.
5. Rinse with distilled water and air dry the slides for another 2 minutes at room temperature.

Tissue Pretreatment Procedure for Mohs Frozen Tissues

1. Subject tissues to epitope retrieval using Bio SB ImmunoDNA Retriever with Citrate (BSB 0020-BSB 0023), ImmunoDNA Retriever with EDTA (BSB 0030-BSB 0033) or Mohs ImmunoDigestor (BSB 0324- 0326) based on antibody pretreatment recommendations. It is recommended that an user choose either a 3-5 minutes heat-induced epitope retrieval (HIER) method using a TintoDetector ImmunoDNA System (BSB 7000) or a pressure cooker (BSB TintoRetriever Pressure Cooker) at 110 °C, or a 1-minute proteolytic induced epitope retrieval (PIER) method (for Cytokeratins using 10% NBF fixed tissues). Morphology is better with 10% NBF fixed tissues, especially when using proteolytic induced epitope retrieval method, however, antigen maybe easier to detect on the 100% acetone fixed tissue.
2. Two different HIER methods may be used:
1. a. TintoDetector ImmunoDNA System
1. Pair theTintoDetector Cap Gap Plus slides (BSB 7006) with Mohs Frozen Tissues face to face and dip slides into a retrieving solution. Make sure the retrieving solution covers the tissues. Insert the TintoDetector Slide handle with the slides inside the TintoDetector incubatorat110° Cand incubate the slides for 3-5 minutes.
2. a. TintoRetriever Pressure Cooker or Equivalent
1. Place tissues/slides in a staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA, and place on trivet or staining dish support in the pressure cooker. Add 1-2 inches of distilled water to the pressure cooker and turn heat to high (110-121° C). Incubate for 5 minutes. Open and immediately transfer slides to room temperature. Cool off for 5 -10 min
3. 3. For PIER, apply Mohs ImmunoDigestor (BSB 0324- 0326) at room temperature for 1 min then wash.

IHC Detection Procedure

1. After HIER or PIER, transfer slides to ImmunoDNA washer and let stand for 1-2 minutes.
2. For manual staining, perform antibody incubation at ambient temperature. For automated staining methods, perform antibody incubation according to instrument manufacturer’s instructions.
3. Wash slides with ImmunoDNA washer or DI water.
4. Continue IHC detection protocol. Wash slides between each step with ImmunoDNA washer solution.