Alteration of Tissue Marking Dyes Depends on Used Chromogen during Immunohistochemistry
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...
Main Authors: | , , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
MDPI AG
2021-04-01
|
Series: | Cells |
Subjects: | |
Online Access: | https://www.mdpi.com/2073-4409/10/4/835 |
id |
doaj-73ffb42179874881b8fe408872bcc2fb |
---|---|
record_format |
Article |
spelling |
doaj-73ffb42179874881b8fe408872bcc2fb2021-04-08T23:00:19ZengMDPI AGCells2073-44092021-04-011083583510.3390/cells10040835Alteration of Tissue Marking Dyes Depends on Used Chromogen during ImmunohistochemistrySelina Kiefer0Julia Huber1Hannah Füllgraf2Kristin Sörensen3Agnes Csanadi4Maren Nicole Stillger5Martin Werner6Hans-Eckart Schaefer7Peter Bronsert8Konrad Aumann9Institute for Surgical Pathology, Medical Center-University of Freiburg, Breisacherstr. 115A, 79106 Freiburg, GermanyInstitute for Surgical Pathology, Medical Center-University of Freiburg, Breisacherstr. 115A, 79106 Freiburg, GermanyInstitute for Surgical Pathology, Medical Center-University of Freiburg, Breisacherstr. 115A, 79106 Freiburg, GermanyInstitute for Surgical Pathology, Medical Center-University of Freiburg, Breisacherstr. 115A, 79106 Freiburg, GermanyInstitute for Surgical Pathology, Medical Center-University of Freiburg, Breisacherstr. 115A, 79106 Freiburg, GermanyInstitute for Surgical Pathology, Medical Center-University of Freiburg, Breisacherstr. 115A, 79106 Freiburg, GermanyInstitute for Surgical Pathology, Medical Center-University of Freiburg, Breisacherstr. 115A, 79106 Freiburg, GermanyInstitute for Surgical Pathology, Medical Center-University of Freiburg, Breisacherstr. 115A, 79106 Freiburg, GermanyInstitute for Surgical Pathology, Medical Center-University of Freiburg, Breisacherstr. 115A, 79106 Freiburg, GermanyInstitute for Surgical Pathology, Medical Center-University of Freiburg, Breisacherstr. 115A, 79106 Freiburg, GermanyPathological 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<sup>®</sup>) 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 H<sub>2</sub>O<sub>2</sub> 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.https://www.mdpi.com/2073-4409/10/4/835immunohistochemistrytissue markingsurgical marginsmargin inkinggross sectionDAB |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Selina Kiefer Julia Huber Hannah Füllgraf Kristin Sörensen Agnes Csanadi Maren Nicole Stillger Martin Werner Hans-Eckart Schaefer Peter Bronsert Konrad Aumann |
spellingShingle |
Selina Kiefer Julia Huber Hannah Füllgraf Kristin Sörensen Agnes Csanadi Maren Nicole Stillger Martin Werner Hans-Eckart Schaefer Peter Bronsert Konrad Aumann Alteration of Tissue Marking Dyes Depends on Used Chromogen during Immunohistochemistry Cells immunohistochemistry tissue marking surgical margins margin inking gross section DAB |
author_facet |
Selina Kiefer Julia Huber Hannah Füllgraf Kristin Sörensen Agnes Csanadi Maren Nicole Stillger Martin Werner Hans-Eckart Schaefer Peter Bronsert Konrad Aumann |
author_sort |
Selina Kiefer |
title |
Alteration of Tissue Marking Dyes Depends on Used Chromogen during Immunohistochemistry |
title_short |
Alteration of Tissue Marking Dyes Depends on Used Chromogen during Immunohistochemistry |
title_full |
Alteration of Tissue Marking Dyes Depends on Used Chromogen during Immunohistochemistry |
title_fullStr |
Alteration of Tissue Marking Dyes Depends on Used Chromogen during Immunohistochemistry |
title_full_unstemmed |
Alteration of Tissue Marking Dyes Depends on Used Chromogen during Immunohistochemistry |
title_sort |
alteration of tissue marking dyes depends on used chromogen during immunohistochemistry |
publisher |
MDPI AG |
series |
Cells |
issn |
2073-4409 |
publishDate |
2021-04-01 |
description |
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<sup>®</sup>) 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 H<sub>2</sub>O<sub>2</sub> 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. |
topic |
immunohistochemistry tissue marking surgical margins margin inking gross section DAB |
url |
https://www.mdpi.com/2073-4409/10/4/835 |
work_keys_str_mv |
AT selinakiefer alterationoftissuemarkingdyesdependsonusedchromogenduringimmunohistochemistry AT juliahuber alterationoftissuemarkingdyesdependsonusedchromogenduringimmunohistochemistry AT hannahfullgraf alterationoftissuemarkingdyesdependsonusedchromogenduringimmunohistochemistry AT kristinsorensen alterationoftissuemarkingdyesdependsonusedchromogenduringimmunohistochemistry AT agnescsanadi alterationoftissuemarkingdyesdependsonusedchromogenduringimmunohistochemistry AT marennicolestillger alterationoftissuemarkingdyesdependsonusedchromogenduringimmunohistochemistry AT martinwerner alterationoftissuemarkingdyesdependsonusedchromogenduringimmunohistochemistry AT hanseckartschaefer alterationoftissuemarkingdyesdependsonusedchromogenduringimmunohistochemistry AT peterbronsert alterationoftissuemarkingdyesdependsonusedchromogenduringimmunohistochemistry AT konradaumann alterationoftissuemarkingdyesdependsonusedchromogenduringimmunohistochemistry |
_version_ |
1721533580610371584 |