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

• Published in: STAR protocols Vol. 1; no. 3; p. 100221
• Main Authors: Geisterfer, Zachary M., Oakey, John, Gatlin, Jesse C.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 18.12.2020; Elsevier
• Subjects: Animals; Biophysics; Cell Biology; Cell Extracts - isolation & purification; Cell isolation; Cell-Free System - metabolism; Cell-Free System - physiology; Cytoplasm - metabolism; Hydrogels - chemistry; Microfluidic Analytical Techniques - methods; Microfluidics - methods; Microscopy; Model Organisms; Oocytes - metabolism; Protocol; Xenopus laevis - metabolism; Model Organisms; Biophysics; Microscopy; Cell isolation; Cell Biology
• DOI: 10.1016/j.xpro.2020.100221
• ISSN: 2666-1667

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). [Display omitted] •Xenopus laevis cell-free extracts can be isolated as discreet volumes typical of cells•Approach provides enhanced control over the shape and position of encapsulated extract•Improved signal-to-noise during fluorescence imaging versus oil-emulsion extract droplets 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.