Abstract
Efficient lysis is critical when analyzing single cells in microfluidic droplets, but existing methods utilize detergents that can interfere with the assays to be performed. We demonstrate robust cell lysis without the use of detergents or other chemicals. In our method, cells are exposed to electric field immediately before encapsulation in droplets, resulting in cell lysis. We characterize lysis efficiency as a function of control parameters and demonstrate compatibility with enzymatic assays by measuring the catalysis of β-glucosidase, an important cellulase used in the conversion of biomass to biofuel. Our method enables assays in microfluidic droplets that are incompatible with detergents.
Highlights
Cellular heterogeneity is important in a variety of biological systems, from providing robustness to evolutionary stresses to enabling effective immune responses against diverse threats.1–3 Because the heterogeneity exists at the level of single cells, studying these systems requires methods for high-throughput single cell analysis
Efficient lysis is critical when analyzing single cells in microfluidic droplets, but existing methods utilize detergents that can interfere with the assays to be performed
Our method enables assays in microfluidic droplets that are incompatible with detergents
Summary
Cellular heterogeneity is important in a variety of biological systems, from providing robustness to evolutionary stresses to enabling effective immune responses against diverse threats. Because the heterogeneity exists at the level of single cells, studying these systems requires methods for high-throughput single cell analysis. Droplet microfluidics breaks through this barrier by allowing single cells to be analyzed using soluble assays, such as enzyme catalysis, detection of secreted products, or presence of unique nucleic acid sequences.. Droplet microfluidics breaks through this barrier by allowing single cells to be analyzed using soluble assays, such as enzyme catalysis, detection of secreted products, or presence of unique nucleic acid sequences.9,10 The devices achieve this using tools for rapidly generating, merging, injecting, and sorting droplets for applications including single-cell sequencing, directed evolution, and drug screening.. Proteases can digest the enzymes necessary for assays, while detergents are difficult to remove from droplets once added and can interfere with important interactions between molecules When using these components to lyse cells, compatible assays must be carefully selected and, even the assay may be influenced by their presence. Detergents commonly used for lysing cells can perturb the stability and activity of enzymes so that measurements performed with detergents often do not agree with the ones performed detergent-free. To enable greater flexibility when choosing assays with which to analyze single cells in microfluidic droplets, new, chemical-free methods are needed for lysing cells
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
