Journal
LAB ON A CHIP
Volume 22, Issue 17, Pages 3258-3267Publisher
ROYAL SOC CHEMISTRY
DOI: 10.1039/d2lc00435f
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Funding
- National Institute of Health [1R33CA223908-01, 5R01GM127714-04]
- National Science Foundation [CBET-1803872]
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Most common cellular analysis methods employ a top-down approach, which destroys the cell and prevents correlation between genomics and functional assays. This study presents a bottom-up approach using single-cell tools, allowing functional phenotyping through observing cytotoxicity and probing underlying biology.
Most common methods of cellular analysis employ the top-down approach (investigating proteomics or genomics directly), thereby destroying the cell, which does not allow the possibility of using the same cell to correlate genomics with functional assays. Herein we describe an approach for single-cell tools that serve as a bottom-up approach. Our technology allows functional phenotyping to be conducted by observing the cytotoxicity of cells and then probe the underlying biology. We have developed a droplet microfluidic device capable of trapping droplets in the array and releasing the droplet of interest selectively using microvalves. Each droplet in the array encapsulates natural killer cells (NK cells) and tumour cells for real-time monitoring of burst kinetics and spatial coordination during killing by single NK cells. Finally, we use the microvalve actuation to selectively release droplets with the desired functional phenotype such as for fast and serial killing of target tumour cells by NK cells. From this perspective, our device allows for investigating first interactions and real-time monitoring of kinetics and later cell recovery on demand for single-cell omic analysis such as single-cell RNA sequencing (scRNA), which to date, is primarily based on in-depth analyses of the entire transcriptome of a relatively low number of cells.
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