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Electromechanical Lysis and Rapid Enzymatic Assay of Droplet-encapsulated Cells
Wednesday, March 10, 2021: 8:40 AM - 9:15 AM
Speaker(s)
Description
Encapsulation of individual tumor cells in water-in-oil droplets supports assays that uncover cell-to-cell variations that drive disease progression and treatment outcomes. Obtaining a distribution of gene expression or enzymatic activity among individual cells can identify subpopulations that are resistant to a chemotherapeutic agent or that are particularly invasive. Droplet microfluidics achieves many of the functions needed for cell analysis through merging, splitting, sorting, and thermal cycling as well as in-droplet mixing. Cell lysis is frequently accomplished by heat, which increases instrument complexity and can denature proteins, or by an added chemical agent, which can interfere with subsequent reactions. Similarly, while merging and mixing readily accomplish dilution, concentration enrichment is advantageous for many applications but is difficult to achieve once droplets are formed. In enzymatic assays, for instance, the reaction rate depends linearly on the concentration of the enzyme, and the limit of detection for the product is also concentration dependent. In this presentation, we describe an electrokinetic method to drive cell lysis and subsequent concentration enrichment (>15-fold) within droplets, thereby achieving increased assay speed and sensitivity. This approach can be applied to many droplets simultaneously and is readily integrated with existing droplet workflows. Using a coupled experimental and computational approach, we discriminate the role of the electric field, osmotic pressure, and shear force in cell lysis and quantify the influence of enrichment on reaction rate. We then leverage this method to determine the distribution in the activity of beta-galactosidase, a marker of cell senescence linked to drug efficacy, among a population of breast cancer cells. Our results demonstrate that electrokinetic manipulation of droplet contents have the potential for broad impact in cell analysis.
Additional Info
Keywords: Please select up to 4 keywords ONLY:
Biotechnology,Enzyme Assays,Sample Preparation
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