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

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

Content Provider	Semantic Scholar
Author	Moon, Hui Sung Je, Kwanghwi Min, Jae-Woong Park, Donghyun Han, Kyung-Yeon Shin, Seung-Ho Park, Woong-Yang Yoo, Chang Eun Kim, Shin-Hyun
Copyright Year	2018
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.
Starting Page	775
Ending Page	784
Page Count	10
File Format	PDF HTM / HTML
Alternate Webpage(s)	http://www.rsc.org/suppdata/c7/lc/c7lc01284e/c7lc01284e5.pdf
PubMed reference number	29423464v1
Alternate Webpage(s)	https://doi.org/10.1039/c7lc01284e
DOI	10.1039/c7lc01284e
Journal	Lab on a chip
Volume Number	18
Issue Number	5
Language	English
Access Restriction	Open
Subject Keyword	Anatomical compartments Base Sequence Bead Dosage Form Biopolymer Sequencing Cell Count Concentrate Dosage Form Instrumentation (attribute) Microfluidics Molecular Profiling One Thousand RNA Sequence Number Transcript
Content Type	Text
Resource Type	Article

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4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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