Assessment of Lung Eosinophils In Situ Using Immunohistological Staining

Christopher D. Nazaroff, William E. LeSuer, Mia Y. Masuda, Grace Pyon, Paige Lacy, Elizabeth A. Jacobsen

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Eosinophils are rare white blood cells that are recruited from circulation to accumulate in the lung in mouse models of allergic respiratory inflammation. In hematoxylin–eosin (HE) stained lungs, eosinophils may be difficult to detect despite their bright eosin staining in the secondary granules. For this reason, antibody-mediated detection of eosinophils is preferable for specific and clearer identification of these cells. Moreover, eosinophils may degranulate, releasing their granule proteins into surrounding tissue, and remnants of cytolysed cells cannot be detected by HE staining. The methods here demonstrate the use of eosinophil-specific anti-mouse antibodies to detect eosinophil granule proteins in formalin-fixed cells both in situ in paraffin-embedded lungs, as well as in cytospin preparations from the lung. These antibody staining techniques enable either colorimetric or fluorescence imaging of eosinophils or their granule proteins with the potential for additional antibodies to be added for detection of multiple molecules.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages237-266
Number of pages30
DOIs
StatePublished - 2021

Publication series

NameMethods in Molecular Biology
Volume2223
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Eosinophil peroxidase
  • Eosinophils
  • Fluorescence
  • Formalin-fixed
  • Granule proteins
  • Immunohistochemistry
  • Lung
  • Major basic protein
  • Staining

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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