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Flow chamber staining modality for real-time inspection of dynamic phenotypes in multiple histological stains

Traditional histological stains, such as hematoxylin-eosin (HE), special stains, and immunofluorescence (IF), have defined myriads of cellular phenotypes and tissue structures in a separate stained section. However, the precise connection of information conveyed by the various stains in the same sec...

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Autores principales: Li, Zhongmin, Muench, Goetz, Goebel, Silvia, Uhland, Kerstin, Wenhart, Clara, Reimann, Andreas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10159194/
https://www.ncbi.nlm.nih.gov/pubmed/37141296
http://dx.doi.org/10.1371/journal.pone.0284444
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author Li, Zhongmin
Muench, Goetz
Goebel, Silvia
Uhland, Kerstin
Wenhart, Clara
Reimann, Andreas
author_facet Li, Zhongmin
Muench, Goetz
Goebel, Silvia
Uhland, Kerstin
Wenhart, Clara
Reimann, Andreas
author_sort Li, Zhongmin
collection PubMed
description Traditional histological stains, such as hematoxylin-eosin (HE), special stains, and immunofluorescence (IF), have defined myriads of cellular phenotypes and tissue structures in a separate stained section. However, the precise connection of information conveyed by the various stains in the same section, which may be important for diagnosis, is absent. Here, we present a new staining modality—Flow chamber stain, which complies with the current staining workflow but possesses newly additional features non-seen in conventional stains, allowing for (1) quickly switching staining modes between destain and restain for multiplex staining in one single section from routinely histological preparation, (2) real-time inspecting and digitally capturing each specific stained phenotype, and (3) efficiently synthesizing graphs containing the tissue multiple-stained components at site-specific regions. Comparisons of its stains with those by the conventional staining fashions using the microscopic images of mouse tissues (lung, heart, liver, kidney, esophagus, and brain), involving stains of HE, Periodic acid–Schiff, Sirius red, and IF for Human IgG, and mouse CD45, hemoglobin, and CD31, showed no major discordance. Repetitive experiments testing on targeted areas of stained sections confirmed the method is reliable with accuracy and high reproducibility. Using the technique, the targets of IF were easily localized and seen structurally in HE- or special-stained sections, and the unknown or suspected components or structures in HE-stained sections were further determined in histological special stains or IF. By the technique, staining processing was videoed and made a backup for off-site pathologists, which facilitates tele-consultation or -education in current digital pathology. Mistakes, which might occur during the staining process, can be immediately found and amended accordingly. With the technique, a single section can provide much more information than the traditional stained counterpart. The staining mode bears great potential to become a common supplementary tool for traditional histopathology.
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spelling pubmed-101591942023-05-05 Flow chamber staining modality for real-time inspection of dynamic phenotypes in multiple histological stains Li, Zhongmin Muench, Goetz Goebel, Silvia Uhland, Kerstin Wenhart, Clara Reimann, Andreas PLoS One Research Article Traditional histological stains, such as hematoxylin-eosin (HE), special stains, and immunofluorescence (IF), have defined myriads of cellular phenotypes and tissue structures in a separate stained section. However, the precise connection of information conveyed by the various stains in the same section, which may be important for diagnosis, is absent. Here, we present a new staining modality—Flow chamber stain, which complies with the current staining workflow but possesses newly additional features non-seen in conventional stains, allowing for (1) quickly switching staining modes between destain and restain for multiplex staining in one single section from routinely histological preparation, (2) real-time inspecting and digitally capturing each specific stained phenotype, and (3) efficiently synthesizing graphs containing the tissue multiple-stained components at site-specific regions. Comparisons of its stains with those by the conventional staining fashions using the microscopic images of mouse tissues (lung, heart, liver, kidney, esophagus, and brain), involving stains of HE, Periodic acid–Schiff, Sirius red, and IF for Human IgG, and mouse CD45, hemoglobin, and CD31, showed no major discordance. Repetitive experiments testing on targeted areas of stained sections confirmed the method is reliable with accuracy and high reproducibility. Using the technique, the targets of IF were easily localized and seen structurally in HE- or special-stained sections, and the unknown or suspected components or structures in HE-stained sections were further determined in histological special stains or IF. By the technique, staining processing was videoed and made a backup for off-site pathologists, which facilitates tele-consultation or -education in current digital pathology. Mistakes, which might occur during the staining process, can be immediately found and amended accordingly. With the technique, a single section can provide much more information than the traditional stained counterpart. The staining mode bears great potential to become a common supplementary tool for traditional histopathology. Public Library of Science 2023-05-04 /pmc/articles/PMC10159194/ /pubmed/37141296 http://dx.doi.org/10.1371/journal.pone.0284444 Text en © 2023 Li et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Li, Zhongmin
Muench, Goetz
Goebel, Silvia
Uhland, Kerstin
Wenhart, Clara
Reimann, Andreas
Flow chamber staining modality for real-time inspection of dynamic phenotypes in multiple histological stains
title Flow chamber staining modality for real-time inspection of dynamic phenotypes in multiple histological stains
title_full Flow chamber staining modality for real-time inspection of dynamic phenotypes in multiple histological stains
title_fullStr Flow chamber staining modality for real-time inspection of dynamic phenotypes in multiple histological stains
title_full_unstemmed Flow chamber staining modality for real-time inspection of dynamic phenotypes in multiple histological stains
title_short Flow chamber staining modality for real-time inspection of dynamic phenotypes in multiple histological stains
title_sort flow chamber staining modality for real-time inspection of dynamic phenotypes in multiple histological stains
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10159194/
https://www.ncbi.nlm.nih.gov/pubmed/37141296
http://dx.doi.org/10.1371/journal.pone.0284444
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