Assessment of Lung Eosinophils In Situ Using Immunohistological Staining.

Autor:	Nazaroff CD; Division of Allergy, Asthma, and Clinical Immunology, Department of Medicine, Mayo Clinic Arizona, Scottsdale, AZ, USA.; Biodesign Institute, School of Molecular Sciences, Arizona State University, Tempe, AZ, USA., LeSuer WE; Division of Allergy, Asthma, and Clinical Immunology, Department of Medicine, Mayo Clinic Arizona, Scottsdale, AZ, USA., Masuda MY; Division of Allergy, Asthma, and Clinical Immunology, Department of Medicine, Mayo Clinic Arizona, Scottsdale, AZ, USA., Pyon G; Division of Allergy, Asthma, and Clinical Immunology, Department of Medicine, Mayo Clinic Arizona, Scottsdale, AZ, USA., Lacy P; Alberta Respiratory Centre (ARC) Research, Department of Medicine, University of Alberta, Edmonton, AB, Canada., Jacobsen EA; Division of Allergy, Asthma, and Clinical Immunology, Department of Medicine, Mayo Clinic Arizona, Scottsdale, AZ, USA. Jacobsen.Elizabeth@Mayo.edu.; Department of Immunology, Mayo Clinic Arizona, Scottsdale, AZ, USA. Jacobsen.Elizabeth@Mayo.edu.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2021; Vol. 2223, pp. 237-266.
DOI:	10.1007/978-1-0716-1001-5_17
Abstrakt:	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.
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu