A protocol for imaging microvilli biogenesis on the surface of cultured porcine kidney epithelial cell monolayers

STAR Protoc. 2021 Dec 10;2(4):100998. doi: 10.1016/j.xpro.2021.100998. eCollection 2021 Dec 17.

Abstract

A key facet of epithelial differentiation is the assembly of actin-based protrusions known as microvilli, which amplify apical membrane surface area for various cell functions. To probe mechanisms of microvillus assembly, we developed a protocol using spinning disk confocal microscopy to directly visualize microvillus biogenesis on the surface of cultured porcine kidney epithelial cell monolayers engineered to express fluorescent proteins. This protocol offers access to the molecular details of individual protrusion growth events at high spatiotemporal resolution. For complete details on the use and execution of this protocol, please refer to Gaeta et al. (2021).

Keywords: Cell Biology; Cell culture; Cell-based Assays; Microscopy.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Cell Culture Techniques
  • Cells, Cultured
  • Epithelial Cells / cytology*
  • Kidney / cytology*
  • Microscopy, Confocal / methods*
  • Microvilli* / chemistry
  • Microvilli* / metabolism
  • Swine