Staining Hypoxic Areas of Frozen and FFPE Tissue Sections with Hypoxyprobe™

Harsh H Oza; Esther Ng; Daniele M Gilkes

doi:10.1007/978-1-0716-3633-6_11

Staining Hypoxic Areas of Frozen and FFPE Tissue Sections with Hypoxyprobe™

Methods Mol Biol. 2024:2755:149-163. doi: 10.1007/978-1-0716-3633-6_11.

Authors

Harsh H Oza¹, Esther Ng², Daniele M Gilkes³

Affiliations

¹ Department of Oncology, The Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins University School of Medicine, Baltimore, MD, USA.
² Department of Biology, The Zanvyl Krieger School of Arts & Sciences, The Johns Hopkins University, Baltimore, MD, USA.
³ Department of Oncology, The Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins University School of Medicine, Baltimore, MD, USA. Dgilkes1@jhu.edu.

PMID: 38319576
DOI: 10.1007/978-1-0716-3633-6_11

Abstract

Hypoxia occurs due to inadequate levels of oxygen in tissue and has been implicated in numerous diseases such as cancer, diabetes, cardiovascular, and neurodegenerative diseases. Hypoxia activates hypoxia-inducible factors (HIF) which mediate the expression of several downstream genes. Within the context of cancer biology, these genes affect cellular processes including metabolism, proliferation, migration, invasion, and metastasis. Pimonidazole hydrochloride (HCl) is an exogenous marker that is reduced and binds to thiols under hypoxic conditions resulting in adducts that can be visualized using antibodies such as Hypoxyprobe™. This chapter describes a method for using Hypoxyprobe™ to detect hypoxic areas in frozen and FFPE mouse samples by immunofluorescence (IF) and immunohistochemistry (IHC) staining.

Keywords: FFPE tissue; Frozen tissue; Hypoxia; Hypoxyprobe™; IF; IHC; Immunofluorescence staining; Immunohistochemistry; Mouse samples; Pimonidazole hydrochloride.

MeSH terms

Animals
Antibodies
Hypoxia*
Mice
Neoplasms*
Oxygen
Staining and Labeling

Substances

Oxygen
Antibodies