Masson Trichrome Staining SOP

Purpose

To provide a standardized method for Masson Trichrome staining to differentiate collagen from muscle and other tissues.

Scope

This SOP applies to all histopathology laboratory personnel performing Masson Trichrome staining.

Responsibilities

• Laboratory Technician: Perform staining procedures
• Quality Manager: Ensure compliance with SOP
• Pathologist: Review stained slides

Materials & equipment

• Masson Trichrome stain kit
• Microscope slides
• Coverslips
• Dehydrating alcohols (Ethanol)
• Xylene
• Distilled water
• Staining dishes

Procedure

1. Prepare tissue sections on slides by deparaffinizing and rehydrating them through graded alcohols to distilled water.
2. Immerse slides in Bouin's solution for 1 hour at 56°C.
3. Wash slides in running tap water for 5 minutes to remove Bouin's solution.
4. Stain slides in Weigert's iron hematoxylin for 5 minutes.
5. Rinse slides in distilled water for 1 minute.
6. Stain slides in the Masson Trichrome stain solution for 15-30 minutes depending on desired intensity.
7. Rinse slides in distilled water for 1 minute.
8. Differentiate slides in 1% acetic acid solution for 1-2 minutes.
9. Dehydrate slides through graded alcohols to xylene.
10. Mount slides with a coverslip using a suitable mounting medium.

Records

• Staining log: Document date, technician name, and batch number
• Quality control records: Include results of control slides

Safety

• Wear appropriate personal protective equipment (PPE) including gloves and lab coat.
• Handle all chemicals according to their safety data sheets (SDS).
• Dispose of waste according to laboratory protocols.