Jump to main content
Jump to site search


Inertial-ordering-assisted droplet microfluidics for high-throughput single-cell RNA-sequencing

Author affiliations

Abstract

Single-cell RNA-seq reveals the cellular heterogeneity inherent in the population of cells, which is very important in many clinical and research applications. Recent advances in droplet microfluidics have achieved the automatic isolation, lysis, and labeling of single cells in droplet compartments without complex instrumentation. However, barcoding errors occurring in the cell encapsulation process because of the multiple-beads-in-droplet and insufficient throughput because of the low concentration of beads for avoiding multiple-beads-in-a-droplet remain important challenges for precise and efficient expression profiling of single cells. In this study, we developed a new droplet-based microfluidic platform that significantly improved the throughput while reducing barcoding errors through deterministic encapsulation of inertially ordered beads. Highly concentrated beads containing oligonucleotide barcodes were spontaneously ordered in a spiral channel by an inertial effect, which were in turn encapsulated in droplets one-by-one, while cells were simultaneously encapsulated in the droplets. The deterministic encapsulation of beads resulted in a high fraction of single-bead-in-a-droplet and rare multiple-beads-in-a-droplet although the bead concentration increased to 1000 μl−1, which diminished barcoding errors and enabled accurate high-throughput barcoding. We successfully validated our device with single-cell RNA-seq. In addition, we found that multiple-beads-in-a-droplet, generated using a normal Drop-Seq device with a high concentration of beads, underestimated transcript numbers and overestimated cell numbers. This accurate high-throughput platform can expand the capability and practicality of Drop-Seq in single-cell analysis.

Graphical abstract: Inertial-ordering-assisted droplet microfluidics for high-throughput single-cell RNA-sequencing

Back to tab navigation

Supplementary files

Publication details

The article was received on 30 Nov 2017, accepted on 31 Jan 2018 and first published on 31 Jan 2018


Article type: Paper
DOI: 10.1039/C7LC01284E
Citation: Lab Chip, 2018, Advance Article
  •   Request permissions

    Inertial-ordering-assisted droplet microfluidics for high-throughput single-cell RNA-sequencing

    H. Moon, K. Je, J. Min, D. Park, K. Han, S. Shin, W. Park, C. E. Yoo and S. Kim, Lab Chip, 2018, Advance Article , DOI: 10.1039/C7LC01284E

Search articles by author

Spotlight

Advertisements