Simplified Drop-seq workflow with minimized bead loss using a bead capture and processing microfluidic chip
Autor: | Esther Amstad, Marjan Biočanin, Johannes Bues, Riccardo Dainese, Bart Deplancke |
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Rok vydání: | 2019 |
Předmět: |
Cell processing
Computer science genetic processes Biomedical Engineering Bioengineering 02 engineering and technology 01 natural sciences Biochemistry Workflow Cell throughput Lab-On-A-Chip Devices Animals Humans natural sciences Droplet microfluidics Sequence Analysis RNA business.industry Drop (liquid) 010401 analytical chemistry General Chemistry 021001 nanoscience & nanotechnology Chip Microspheres 0104 chemical sciences Drosophila melanogaster HEK293 Cells Microfluidic chip Single-Cell Analysis 0210 nano-technology business Computer hardware |
Zdroj: | Lab on a Chip. 19:1610-1620 |
ISSN: | 1473-0189 1473-0197 |
Popis: | 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. |
Databáze: | OpenAIRE |
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