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Ion Concentration Polarization Mediated In-droplet Cell Lysis and Enhancement of Beta-galactosidase Enzymatic Assay
Monday, March 08, 2021: 8:30 AM - 6:00 PM
Speaker(s)
Description
Droplet microfluidics has carved a niche for itself in the world of bioanalytics. The versatility of droplet manipulation allows for the development of droplet-based bioassays and single-cell analysis techniques. While customized droplet generation, splitting, merging, and sorting have been well researched and optimized, in-droplet manipulation still lies in the developmental domain. Our group recently described in-droplet manipulation of concentration distribution and cation exchange by the electrokinetic phenomenon of ion concentration polarization (ICP). We showed that the ionic contents of droplets positioned between two cation selective membranes were polarized under a voltage bias, leading to the enrichment of a fluorescent analyte. In this presentation, we report a method that leverages ICP to achieve in-droplet cell lysis and enhanced beta-galactosidase (beta-gal) enzymatic assay sensitivity.
The study encompasses three main objectives. First, an integrated device is designed for monodisperse nanoliter-scale droplet generation and in-droplet ICP. Second, ICP-mediated in-droplet lysis of encapsulated MDA-MB-231 cells (mammalian adenocarcinoma) in diluted DPBS buffer (Dulbecco’s phosphate buffered saline) is qualitatively demonstrated. Third, we present the quantitative assessment of enhanced in-droplet beta-gal enzymatic assay as a result of analyte enrichment mediated by ICP.
We believe that the present study, centered on ICP and its applications, will allow for the development of high throughput, rapid, and sensitive on-chip single-cell enzymatic assays. An on-chip enhanced beta-gal assay prototype as studied, will therefore be beneficial towards a better understanding of aging and cell apoptosis on the oncological front.
The authors gratefully acknowledge financial support from NSF under award no. 1849109.
Additional Info
Keywords: Please select up to 4 keywords ONLY:
Enzyme Assays,Membrane
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