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Novel Chromogens for Immunohistochemistry in Spatial Biology
Version 1
: Received: 24 April 2024 / Approved: 25 April 2024 / Online: 25 April 2024 (14:23:11 CEST)
A peer-reviewed article of this Preprint also exists.
Gupta, B.; Yang, G.; Key, M. Novel Chromogens for Immunohistochemistry in Spatial Biology. Cells 2024, 13, 936. Gupta, B.; Yang, G.; Key, M. Novel Chromogens for Immunohistochemistry in Spatial Biology. Cells 2024, 13, 936.
Abstract
Spatial relations between tumor cells and host-infiltrating cells are increasingly important in both basic science and clinical research. In this study we have tested the feasibility of using standard methods of immunohistochemistry (IHC) in a multiplex staining system using a newly developed set of chromogenic substrates for the peroxidase and alkaline phosphatase enzymes. Using this approach we have developed a set of chromogens characterized by 1) providing fine cellular detail, 2) non-overlapping spectral profiles, 3) absence of interactions between chromogens, 4) stable upon storage, and 5) compatible with current standard immunohistochemistry practices. When viewed microscopically under brightfield illumination the chromogens yielded the following colors; red, black, blue, yellow, brown, and green. By selecting compatible color combinations we have shown feasibility for four-color multiplex staining. Depending on the particular type of analysis being performed, visual analysis, without the aid of computer assisted image analysis, was sufficient to differentiate up to four different markers.
Keywords
Immunohistochemistry; chromogens; brightfield; spatial biology
Subject
Biology and Life Sciences, Cell and Developmental Biology
Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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