Microfluidic encapsulation of Xenopus laevis cell-free extracts using hydrogel photolithography

STAR Protoc. 2020 Dec 11;1(3):100221. doi: 10.1016/j.xpro.2020.100221. eCollection 2020 Dec 18.

Abstract

Cell-free extract derived from the eggs of the African clawed frog Xenopus laevis is a well-established model system that has been used historically in bulk aliquots. Here, we describe a microfluidic approach for isolating discrete, biologically relevant volumes of cell-free extract, with more expansive and precise control of extract shape compared with extract-oil emulsions. This approach is useful for investigating the mechanics of intracellular processes affected by cell geometry or cytoplasmic volume, including organelle scaling and positioning mechanisms. For complete details on the use and execution of this protocol, please refer to Geisterfer et al. (2020).

Keywords: Biophysics; Cell Biology; Cell isolation; Microscopy; Model Organisms.

Publication types

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

MeSH terms

  • Animals
  • Cell Extracts / isolation & purification*
  • Cell-Free System / metabolism
  • Cell-Free System / physiology
  • Cytoplasm / metabolism
  • Hydrogels / chemistry
  • Microfluidic Analytical Techniques / methods*
  • Microfluidics / methods*
  • Oocytes / metabolism
  • Xenopus laevis / metabolism

Substances

  • Cell Extracts
  • Hydrogels