Cargando…

Blood identified and quantified in formalin fixed paraffin embedded lung sections using eosin fluorescence

Eosin Y is a common stain in histology. Although usually used for colourimetric imaging where the dye is used to stain pink/red a range of structures in the tissue, Eosin Y is also a fluorochrome, and has been used in this manner for decades. In this study our aim was to investigate the fluorescence...

Descripción completa

Detalles Bibliográficos
Autores principales: Francis, Robert J., Ferguson, Deborah, Kempster, Sarah, Hall, Joanna, Berry, Neil, MacLellan-Gibson, Kirsty
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9406261/
https://www.ncbi.nlm.nih.gov/pubmed/36006466
http://dx.doi.org/10.1007/s00418-022-02130-z
_version_ 1784774077588701184
author Francis, Robert J.
Ferguson, Deborah
Kempster, Sarah
Hall, Joanna
Berry, Neil
MacLellan-Gibson, Kirsty
author_facet Francis, Robert J.
Ferguson, Deborah
Kempster, Sarah
Hall, Joanna
Berry, Neil
MacLellan-Gibson, Kirsty
author_sort Francis, Robert J.
collection PubMed
description Eosin Y is a common stain in histology. Although usually used for colourimetric imaging where the dye is used to stain pink/red a range of structures in the tissue, Eosin Y is also a fluorochrome, and has been used in this manner for decades. In this study our aim was to investigate the fluorescence properties of the dye to enable quantification of structures within formalin-fixed paraffin-embedded (FFPE) tissue sections. To do this, FFPE sections of hamster tissue were prepared with haematoxylin and eosin Y dyes. Spectral detection on a confocal laser scanning microscope was used to obtain the fluorescence emission spectra of the eosin Y under blue light. This showed clear spectral differences between the red blood cells and congealed blood, compared to the rest of the section. The spectra were so distinct that it was possible to discern these in fluorescence and multi-photon microscopy. An image analysis algorithm was used to quantify the red blood cells. These analyses could have broad applications in histopathology where differentiation is required, such as the analysis of clotting disorders to haemorrhage or damage from infectious disease. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00418-022-02130-z.
format Online
Article
Text
id pubmed-9406261
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Springer Berlin Heidelberg
record_format MEDLINE/PubMed
spelling pubmed-94062612022-08-26 Blood identified and quantified in formalin fixed paraffin embedded lung sections using eosin fluorescence Francis, Robert J. Ferguson, Deborah Kempster, Sarah Hall, Joanna Berry, Neil MacLellan-Gibson, Kirsty Histochem Cell Biol Short Communication Eosin Y is a common stain in histology. Although usually used for colourimetric imaging where the dye is used to stain pink/red a range of structures in the tissue, Eosin Y is also a fluorochrome, and has been used in this manner for decades. In this study our aim was to investigate the fluorescence properties of the dye to enable quantification of structures within formalin-fixed paraffin-embedded (FFPE) tissue sections. To do this, FFPE sections of hamster tissue were prepared with haematoxylin and eosin Y dyes. Spectral detection on a confocal laser scanning microscope was used to obtain the fluorescence emission spectra of the eosin Y under blue light. This showed clear spectral differences between the red blood cells and congealed blood, compared to the rest of the section. The spectra were so distinct that it was possible to discern these in fluorescence and multi-photon microscopy. An image analysis algorithm was used to quantify the red blood cells. These analyses could have broad applications in histopathology where differentiation is required, such as the analysis of clotting disorders to haemorrhage or damage from infectious disease. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00418-022-02130-z. Springer Berlin Heidelberg 2022-08-25 2022 /pmc/articles/PMC9406261/ /pubmed/36006466 http://dx.doi.org/10.1007/s00418-022-02130-z Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Short Communication
Francis, Robert J.
Ferguson, Deborah
Kempster, Sarah
Hall, Joanna
Berry, Neil
MacLellan-Gibson, Kirsty
Blood identified and quantified in formalin fixed paraffin embedded lung sections using eosin fluorescence
title Blood identified and quantified in formalin fixed paraffin embedded lung sections using eosin fluorescence
title_full Blood identified and quantified in formalin fixed paraffin embedded lung sections using eosin fluorescence
title_fullStr Blood identified and quantified in formalin fixed paraffin embedded lung sections using eosin fluorescence
title_full_unstemmed Blood identified and quantified in formalin fixed paraffin embedded lung sections using eosin fluorescence
title_short Blood identified and quantified in formalin fixed paraffin embedded lung sections using eosin fluorescence
title_sort blood identified and quantified in formalin fixed paraffin embedded lung sections using eosin fluorescence
topic Short Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9406261/
https://www.ncbi.nlm.nih.gov/pubmed/36006466
http://dx.doi.org/10.1007/s00418-022-02130-z
work_keys_str_mv AT francisrobertj bloodidentifiedandquantifiedinformalinfixedparaffinembeddedlungsectionsusingeosinfluorescence
AT fergusondeborah bloodidentifiedandquantifiedinformalinfixedparaffinembeddedlungsectionsusingeosinfluorescence
AT kempstersarah bloodidentifiedandquantifiedinformalinfixedparaffinembeddedlungsectionsusingeosinfluorescence
AT halljoanna bloodidentifiedandquantifiedinformalinfixedparaffinembeddedlungsectionsusingeosinfluorescence
AT berryneil bloodidentifiedandquantifiedinformalinfixedparaffinembeddedlungsectionsusingeosinfluorescence
AT maclellangibsonkirsty bloodidentifiedandquantifiedinformalinfixedparaffinembeddedlungsectionsusingeosinfluorescence