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Simplified Drop-seq workflow with minimized bead loss using a bead capture and processing microfluidic chip.

Lab on a chip (2019-03-29)
Marjan Biočanin, Johannes Bues, Riccardo Dainese, Esther Amstad, Bart Deplancke
ABSTRACT

Single-cell RNA-sequencing (scRNA-seq) has revolutionized biomedical research by enabling the in-depth analysis of cell-to-cell heterogeneity of tissues with unprecedented resolution. One of the catalyzing technologies is single cell droplet microfluidics, which has massively increased the overall cell throughput, routinely allowing the analysis of thousands of cells per experiment at a relatively low cost. Among several existing droplet-based approaches, the Drop-seq platform has emerged as one of the most widely used systems. Yet, this has surprisingly not incentivized major refinements of the method, thus restricting any lab implementation to the original Drop-seq setup, which is known to suffer from up to 80% bead loss during the process. In this study, we present a systematic re-engineering and optimization of Drop-seq: first, we re-designed the original dropleting device to be compatible with both air-pressure systems and syringe pumps, thus increasing the overall flexibility of the platform. Second, we devised an accompanying chip for post-encapsulation bead processing, which simplifies and massively increases Drop-seq's cell processing efficiency. Taken together, the presented optimization efforts result in a more flexible and efficient Drop-seq version.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Trichloro(1H,1H,2H,2H-perfluorooctyl)silane, 97%
SAFC
Trypsin-EDTA Solution 10X