Multispectral Staining and Analysis of Extracellular Matrix

Carine M Renner; Mike R Visetsouk; Pamela K Kreeger; Kristyn S Masters

doi:10.1007/978-1-0716-1956-8_6

Multispectral Staining and Analysis of Extracellular Matrix

Methods Mol Biol. 2022:2424:105-119. doi: 10.1007/978-1-0716-1956-8_6.

Authors

Carine M Renner¹, Mike R Visetsouk¹, Pamela K Kreeger^{1

2}, Kristyn S Masters^{3

4}

Affiliations

¹ Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA.
² Department of Cell and Regenerative Biology, Department of Obstetrics and Gynecology, and University of Wisconsin Carbone Cancer Center, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA.
³ Department of Biomedical Engineering and Department of Materials Science and Engineering, University of Wisconsin-Madison, Madison, WI, USA. kmasters@wisc.edu.
⁴ Departments of Medicine, University of Wisconsin Carbone Cancer Center, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA. kmasters@wisc.edu.

PMID: 34918289
DOI: 10.1007/978-1-0716-1956-8_6

Abstract

Multiplexed immunofluorescent (IF) techniques enable the detection of multiple antigens within the same sample and are therefore useful in situations where samples are rare or small in size. Similar to standard IF, multiplexed IF yields information on both the location and relative amount of detected antigens. While this method has been used primarily to detail cell phenotypes, we have recently adapted it to profile the extracellular matrix (ECM), which provides technical challenges due to autofluorescence and spatial overlap. This chapter details the planning, execution, optimization, and troubleshooting to use multiplexed IF to profile the ECM of human fallopian tube tissue.

Keywords: Ovarian cancer; STIC; Serous tubal intra-epithelial carcinoma.

MeSH terms

Carcinoma in Situ
Cystadenocarcinoma, Serous
Extracellular Matrix*
Fallopian Tube Neoplasms
Fallopian Tubes
Female
Humans
Ovarian Neoplasms
Staining and Labeling