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.