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Massively parallel encapsulation of single cells with structured microparticles and secretion-based flow sorting

biorxiv. 2020; 
Joseph de Rutte,  Robert Dimatteo,  Mark van Zee,  Robert Damoiseaux,  Dino Di Carlo
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Catalog Peptides A PEG phase comprised of 17.5% w/w 4-arm PEG-Norbornene (Sigma), 2% w/w LAP (Lithium phenyl-2,4,6-trimethylbenzoylphosphinate,sigma), and 0.5 mg/ml Biotin-PEG-thiol (5000 MW, Nanocs) in Phosphate Buffered Saline (PBS, pH7.2) was co-injected with a dextran phase comprised of 20% w/w 40 kDa dextran (Sigma), 1.3% w/w DTT (dithiothreitol, Sigma), and 5 mM RGD peptide (Ac-RGDSPGERCGNH2, Genscript) in PBS at a rate of 8 µL/min and 2.67 µL/min, respectively using syringe pumps (Harvard Apparatus PHD 2000). Get A Quote

摘要

Techniques to analyze and sort single cells based on secreted products have the potential to transform our understanding of cellular biology as well as accelerate the development of next generation cell and antibody therapies. However, secretions are rapidly transported away from cells, such that specialized equipment and expertise has been required to compartmentalize cells and capture their secretions. Herein we demonstrate the use of cavity-containing hydrogel microparticles to perform functional single-cell secretion analysis and sorting using only commonly accessible lab infrastructure. These microparticles act as a solid support which facilitates cell attachment, templates formation of uniform aqueous com... More

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