Imaging and quantitative analysis of integrin-dependent cell-matrix adhesions

STAR Protoc. 2023 Sep 15;4(3):102473. doi: 10.1016/j.xpro.2023.102473. Epub 2023 Aug 23.

Abstract

Integrin-dependent cell-extracellular matrix adhesion is essential for wound healing, embryonic development, immunity, and tissue organization. Here, we present a protocol for the imaging and quantitative analysis of integrin-dependent cell-matrix adhesions. We describe steps for cell culture; virus preparation; lentiviral transduction; imaging with widefield, confocal, and total internal reflection fluorescence microscopy; and using a script for their quantitative analysis. We then detail procedures for analyzing adhesion dynamics by live-cell imaging and fluorescence recovery after photobleaching (FRAP). For complete details on the use and execution of this protocol, please refer to Margadant et al. (2012),1 van der Bijl et al. (2020),2 Amado-Azevedo et al. (2021).3.

Keywords: Cell Biology; Cell Culture; Microscopy.

MeSH terms

  • Cell Culture Techniques*
  • Cell-Matrix Junctions
  • Embryonic Development
  • Female
  • Humans
  • Integrins
  • Microscopy*
  • Pregnancy

Substances

  • Integrins