Concurrent Viewing of H&E and Multiplex Immunohistochemistry in Clinical Specimens

<b>Background/Objectives:</b> Performing hematoxylin and eosin (H&E) staining and immunohistochemistry (IHC) on the same specimen slide provides advantages that include specimen conservation and the ability to combine the H&E context with biomarker expression at the individual ce...

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Main Authors: Larry E. Morrison, Tania M. Larrinaga, Brian D. Kelly, Mark R. Lefever, Rachel C. Beck, Daniel R. Bauer
Format: Article
Language:English
Published: MDPI AG 2025-01-01
Series:Diagnostics
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Online Access:https://www.mdpi.com/2075-4418/15/2/164
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author Larry E. Morrison
Tania M. Larrinaga
Brian D. Kelly
Mark R. Lefever
Rachel C. Beck
Daniel R. Bauer
author_facet Larry E. Morrison
Tania M. Larrinaga
Brian D. Kelly
Mark R. Lefever
Rachel C. Beck
Daniel R. Bauer
author_sort Larry E. Morrison
collection DOAJ
description <b>Background/Objectives:</b> Performing hematoxylin and eosin (H&E) staining and immunohistochemistry (IHC) on the same specimen slide provides advantages that include specimen conservation and the ability to combine the H&E context with biomarker expression at the individual cell level. We previously used invisible deposited chromogens and dual-camera imaging, including monochrome and color cameras, to implement simultaneous H&E and IHC. Using this approach, conventional H&E staining could be simultaneously viewed in color on a computer monitor alongside a monochrome video of the invisible IHC staining, while manually scanning the specimen. <b>Methods:</b> We have now simplified the microscope system to a single camera and increased the IHC multiplexing to four biomarkers using translational assays. The color camera used in this approach also enabled multispectral imaging, similar to monochrome cameras. <b>Results:</b> Application is made to several clinically relevant specimens, including breast cancer (HER2, ER, and PR), prostate cancer (PSMA, P504S, basal cell, and CD8), Hodgkin’s lymphoma (CD15 and CD30), and melanoma (LAG3). Additionally, invisible chromogenic IHC was combined with conventional DAB IHC to present a multiplex IHC assay with unobscured DAB staining, suitable for visual interrogation. <b>Conclusions:</b> Simultaneous staining and detection, as described here, provides the pathologist a means to evaluate complex multiplexed assays, while seated at the microscope, with the added multispectral imaging capability to support digital pathology and artificial intelligence workflows of the future.
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spelling doaj-art-7c92bd73b1f94cc7833cf3b18c972aeb2025-01-24T13:28:57ZengMDPI AGDiagnostics2075-44182025-01-0115216410.3390/diagnostics15020164Concurrent Viewing of H&E and Multiplex Immunohistochemistry in Clinical SpecimensLarry E. Morrison0Tania M. Larrinaga1Brian D. Kelly2Mark R. Lefever3Rachel C. Beck4Daniel R. Bauer5Roche Diagnostics Solutions (Ventana Medical Systems, Inc.), 1910 E. Innovation Park Dr., Tucson, AZ 85755, USARoche Diagnostics Solutions (Ventana Medical Systems, Inc.), 1910 E. Innovation Park Dr., Tucson, AZ 85755, USARoche Diagnostics Solutions (Ventana Medical Systems, Inc.), 1910 E. Innovation Park Dr., Tucson, AZ 85755, USARoche Diagnostics Solutions (Ventana Medical Systems, Inc.), 1910 E. Innovation Park Dr., Tucson, AZ 85755, USARoche Diagnostics Solutions (Ventana Medical Systems, Inc.), 1910 E. Innovation Park Dr., Tucson, AZ 85755, USARoche Diagnostics Solutions (Ventana Medical Systems, Inc.), 1910 E. Innovation Park Dr., Tucson, AZ 85755, USA<b>Background/Objectives:</b> Performing hematoxylin and eosin (H&E) staining and immunohistochemistry (IHC) on the same specimen slide provides advantages that include specimen conservation and the ability to combine the H&E context with biomarker expression at the individual cell level. We previously used invisible deposited chromogens and dual-camera imaging, including monochrome and color cameras, to implement simultaneous H&E and IHC. Using this approach, conventional H&E staining could be simultaneously viewed in color on a computer monitor alongside a monochrome video of the invisible IHC staining, while manually scanning the specimen. <b>Methods:</b> We have now simplified the microscope system to a single camera and increased the IHC multiplexing to four biomarkers using translational assays. The color camera used in this approach also enabled multispectral imaging, similar to monochrome cameras. <b>Results:</b> Application is made to several clinically relevant specimens, including breast cancer (HER2, ER, and PR), prostate cancer (PSMA, P504S, basal cell, and CD8), Hodgkin’s lymphoma (CD15 and CD30), and melanoma (LAG3). Additionally, invisible chromogenic IHC was combined with conventional DAB IHC to present a multiplex IHC assay with unobscured DAB staining, suitable for visual interrogation. <b>Conclusions:</b> Simultaneous staining and detection, as described here, provides the pathologist a means to evaluate complex multiplexed assays, while seated at the microscope, with the added multispectral imaging capability to support digital pathology and artificial intelligence workflows of the future.https://www.mdpi.com/2075-4418/15/2/164immunohistochemistry (IHC)multiplexinghematoxylin and eosin (H&E)chromogensbrightfield microscopymultispectral imaging
spellingShingle Larry E. Morrison
Tania M. Larrinaga
Brian D. Kelly
Mark R. Lefever
Rachel C. Beck
Daniel R. Bauer
Concurrent Viewing of H&E and Multiplex Immunohistochemistry in Clinical Specimens
Diagnostics
immunohistochemistry (IHC)
multiplexing
hematoxylin and eosin (H&E)
chromogens
brightfield microscopy
multispectral imaging
title Concurrent Viewing of H&E and Multiplex Immunohistochemistry in Clinical Specimens
title_full Concurrent Viewing of H&E and Multiplex Immunohistochemistry in Clinical Specimens
title_fullStr Concurrent Viewing of H&E and Multiplex Immunohistochemistry in Clinical Specimens
title_full_unstemmed Concurrent Viewing of H&E and Multiplex Immunohistochemistry in Clinical Specimens
title_short Concurrent Viewing of H&E and Multiplex Immunohistochemistry in Clinical Specimens
title_sort concurrent viewing of h e and multiplex immunohistochemistry in clinical specimens
topic immunohistochemistry (IHC)
multiplexing
hematoxylin and eosin (H&E)
chromogens
brightfield microscopy
multispectral imaging
url https://www.mdpi.com/2075-4418/15/2/164
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AT markrlefever concurrentviewingofheandmultipleximmunohistochemistryinclinicalspecimens
AT rachelcbeck concurrentviewingofheandmultipleximmunohistochemistryinclinicalspecimens
AT danielrbauer concurrentviewingofheandmultipleximmunohistochemistryinclinicalspecimens