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Alteration of Tissue Marking Dyes Depends on Used Chromogen during Immunohistochemistry

Authors :
Selina Kiefer
Julia Huber
Hannah Füllgraf
Kristin Sörensen
Agnes Csanadi
Maren Nicole Stillger
Martin Werner
Hans-Eckart Schaefer
Peter Bronsert
Konrad Aumann
Source :
Cells, Vol 10, Iss 4, p 835 (2021)
Publication Year :
2021
Publisher :
MDPI AG, 2021.

Abstract

Pathological biopsy protocols require tissue marking dye (TMD) for orientation. In some cases (e.g., close margin), additional immunohistochemical analyses can be necessary. Therefore, the correlation between the applied TMD during macroscopy and the examined TMD during microscopy is crucial for the correct orientation, the residual tumour status and the subsequent therapeutic regime. In this context, our group observed colour changes during routine immunohistochemistry. Tissue specimens were marked with various TMD and processed by two different methods. TMD (blue, red, black, yellow and green) obtained from three different providers (A, B and C, and Whiteout/Tipp-Ex®) were used. Immunohistochemistry was performed manually via stepwise omission of reagents to identify the colour changing mechanism. Blue colour from provider A changed during immunohistochemistry into black, when 3,3′-Diaminobenzidine-tetrahydrochloride-dihydrate (DAB) and H2O2 was applied as an immunoperoxidase-based terminal colour signal. No other applied reagents, nor tissue texture or processing showed any influence on the colour. The remaining colours from provider A and the other colours did not show any changes during immunohistochemistry. Our results demonstrate an interesting and important pitfall in routine immunohistochemistry-based diagnostics that pathologists should be aware of. Furthermore, the chemical rationale behind the observed misleading colour change is discussed.

Details

Language :
English
ISSN :
20734409 and 42179874
Volume :
10
Issue :
4
Database :
Directory of Open Access Journals
Journal :
Cells
Publication Type :
Academic Journal
Accession number :
edsdoj.73ffb42179874881b8fe408872bcc2fb
Document Type :
article
Full Text :
https://doi.org/10.3390/cells10040835