Issue 9, 2019

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

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.

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

Supplementary files

Article information

Article type
Paper
Submitted
04 yan 2019
Accepted
09 fev 2019
First published
28 mar 2019
This article is Open Access
Creative Commons BY-NC license

Lab Chip, 2019,19, 1610-1620

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

M. Biočanin, J. Bues, R. Dainese, E. Amstad and B. Deplancke, Lab Chip, 2019, 19, 1610 DOI: 10.1039/C9LC00014C

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