Incomplete Staining Artifact: A Confounding Frozen Section Pathology Artifact Encountered During Mohs Micrographic...

Area of Science:

• Dermatopathology
• Histotechnology
• Mohs Micrographic Surgery

Background:

• Frozen section pathology artifacts can complicate histological interpretation during Mohs Micrographic Surgery (MMS).
• A specific amorphous, eosinophilic artifact obscuring cellular architecture was repeatedly observed.
• This artifact was suspected to be related to incomplete tissue staining, potentially due to hematoxylin and acetone interactions.

Purpose of the Study:

• To describe a unique incomplete staining frozen section pathology artifact encountered in Mohs Micrographic Surgery.
• To identify the cause of the artifact and implement a protocol adjustment to resolve it.

Main Methods:

• Histological interpretation of frozen sections during Mohs Micrographic Surgery.
• Analysis of staining procedures, specifically focusing on the interaction between hematoxylin and acetone.
• Modification of the SLS stain line protocol by introducing a 15-second water rinse between acetone and hematoxylin steps.
• Comparison of pre- and post-protocol adjustment slide quality.

Main Results:

• The amorphous, eosinophilic artifact was consistently observed across multiple Mohs Micrographic Surgery laboratories.
• The adjusted SLS staining protocol, including the water rinse, successfully resolved the incomplete staining artifact.
• Histological interpretation was no longer obscured by the artifact in the modified protocol.

Conclusions:

• Incomplete tissue staining, potentially due to hematoxylin and acetone interactions, is a identifiable frozen section artifact in Mohs Micrographic Surgery.
• A simple modification to the staining protocol, specifically adding a water rinse, effectively eliminates this artifact.
• Standardizing staining protocols is crucial for accurate intraoperative histological interpretation in Mohs Micrographic Surgery.