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. 2022:2413:37-43.
doi: 10.1007/978-1-0716-1896-7_5.

Immunofluorescence-Based Method to Assess Cancer Biomarker in the Hypoxic Region of the Tumor

Leslimar Rios-Colon  1   2 , Susy Kim  1 , Yixin Su  1 , Deepak Kumar  2 , Gagan Deep  3   4
Affiliations

Immunofluorescence-Based Method to Assess Cancer Biomarker in the Hypoxic Region of the Tumor

Leslimar Rios-Colon et al. Methods Mol Biol. 2022.

Abstract

The development of novel imaging technologies allows the analysis of the expression and spatial distribution of multiple markers simultaneously, providing necessary information about a cellular identity and the surrounding microenvironment. This chapter describes the utilization of immunofluorescence to identify such biomarkers in fixed tissue from prostate cancer (PCa) xenografts. One such marker detectable by immunofluorescence is pimonidazole, which has been utilized to locate areas of low oxygen (hypoxia). Pimonidazole, in combination with other biomarkers, could be utilized to identify "niches" in the microenvironment harboring more aggressive cells both within and outside hypoxic areas. Specifically, we describe the method to use pimonidazole for the identification of hypoxic regions in PCa xenograft tumors along with CPT1A (carnitine palmitoyltransferase 1A) expression, an indicator of β-oxidation. This approach could be useful to characterize various biomarkers in the complex hypoxic tumor microenvironment.

Keywords: Biomarker; Cancer; Hypoxia; Immunofluorescence; Pimonidazole.

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Figures

Fig. 1
Fig. 1
Assessing the expression of a biomarker in hypoxic region of the prostate tumors by pimonidazole staining. Male nude mice bearing 22Rv1 xenografts were injected pimonidazole (IP) 2 h before sacrifice. Next, tumors were collected in OCT, cryosectioned, and processed for confocal analyses using DAPI (blue), pimonidazole-FITC (green), and Alexa Fluor 555 (red) for CPT1A. Representative images (single and merged) are shown at 600×
See this image and copyright information in PMC

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