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Alteration of Tissue Marking Dyes Depends on Used Chromogen during Immunohistochemistry

1
Institute for Surgical Pathology, Medical Center-University of Freiburg, Breisacherstr. 115A, 79106 Freiburg im Breisgau, Germany
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Comprehensive Cancer Center Freiburg, Medical Center-University of Freiburg, 79106 Freiburg im Breisgau, Germany
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Faculty of Biology, University of Freiburg, 79106 Freiburg im Breisgau, Germany
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Institute of Molecular Medicine and Cell Research, Medical Center, University of Freiburg, 79106 Freiburg im Breisgau, Germany
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Center for Pathology Kempten Allgäu-Klinikverbund Allgäu, 87439 Kempten, Germany
*
Author to whom correspondence should be addressed.
Academic Editor: Alexander E. Kalyuzhny
Cells 2021, 10(4), 835; https://doi.org/10.3390/cells10040835
Received: 9 March 2021 / Revised: 1 April 2021 / Accepted: 6 April 2021 / Published: 8 April 2021
(This article belongs to the Special Issue Immuno-Metabolic Crosstalk in Oncogenesis)
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. View Full-Text
Keywords: immunohistochemistry; tissue marking; surgical margins; margin inking; gross section; DAB; Diaminobenzidine-tetrahydrochloride-dihydrate immunohistochemistry; tissue marking; surgical margins; margin inking; gross section; DAB; Diaminobenzidine-tetrahydrochloride-dihydrate
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MDPI and ACS Style

Kiefer, S.; Huber, J.; Füllgraf, H.; Sörensen, K.; Csanadi, A.; Stillger, M.N.; Werner, M.; Schaefer, H.-E.; Bronsert, P.; Aumann, K. Alteration of Tissue Marking Dyes Depends on Used Chromogen during Immunohistochemistry. Cells 2021, 10, 835. https://doi.org/10.3390/cells10040835

AMA Style

Kiefer S, Huber J, Füllgraf H, Sörensen K, Csanadi A, Stillger MN, Werner M, Schaefer H-E, Bronsert P, Aumann K. Alteration of Tissue Marking Dyes Depends on Used Chromogen during Immunohistochemistry. Cells. 2021; 10(4):835. https://doi.org/10.3390/cells10040835

Chicago/Turabian Style

Kiefer, Selina; Huber, Julia; Füllgraf, Hannah; Sörensen, Kristin; Csanadi, Agnes; Stillger, Maren N.; Werner, Martin; Schaefer, Hans-Eckart; Bronsert, Peter; Aumann, Konrad. 2021. "Alteration of Tissue Marking Dyes Depends on Used Chromogen during Immunohistochemistry" Cells 10, no. 4: 835. https://doi.org/10.3390/cells10040835

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