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

Robert J Francis1, Deborah Ferguson2, Sarah Kempster3

  • 1Biological Imaging Group, National Institute for Biological Standards and Control, Blanche Lane, South Mimms, EN6 3QG, UK.

Histochemistry and Cell Biology
|August 25, 2022
PubMed
Summary

Eosin Y, a common histology stain, can be used as a fluorescent dye. This study quanties red blood cells in tissue sections using Eosin Y fluorescence, aiding histopathology analysis.

Keywords:
EosinErythrocytesFFPEH&ESectionSpectral

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Area of Science:

  • Histology
  • Fluorescence Microscopy
  • Biomedical Imaging

Background:

  • Eosin Y is a widely used histological stain for colorimetric imaging.
  • Beyond its colorimetric use, Eosin Y possesses inherent fluorescence properties.
  • Its potential for quantitative analysis in fluorescence microscopy remains underexplored.

Purpose of the Study:

  • To investigate the fluorescence characteristics of Eosin Y in formalin-fixed paraffin-embedded (FFPE) tissue sections.
  • To develop a method for quantifying specific tissue structures using Eosin Y fluorescence.
  • To explore the utility of Eosin Y fluorescence in histopathological analysis.

Main Methods:

  • Prepared FFPE hamster tissue sections stained with haematoxylin and Eosin Y.
  • Acquired fluorescence emission spectra of Eosin Y using confocal laser scanning microscopy under blue light.
  • Utilized an image analysis algorithm for quantification of red blood cells.

Main Results:

  • Distinct fluorescence spectral differences were observed between red blood cells/congealed blood and other tissue components.
  • These spectral distinctions enabled clear visualization in fluorescence and multi-photon microscopy.
  • An image analysis algorithm successfully quantified red blood cells based on Eosin Y fluorescence.

Conclusions:

  • Eosin Y can be effectively utilized as a fluorochrome for quantitative analysis in FFPE tissues.
  • The distinct fluorescence of Eosin Y allows for the differentiation and quantification of specific structures like red blood cells.
  • This approach has potential applications in histopathology for analyzing conditions involving blood, such as clotting disorders or hemorrhage.