Single-cell isolation by a modular single-cell pipette for RNA-sequencing.

TitleSingle-cell isolation by a modular single-cell pipette for RNA-sequencing.
Publication TypeJournal Article
Year of Publication2016
AuthorsZhang, K, Gao, M, Chong, Z, Li, Y, Han, X, Chen, R, Qin, L
JournalLab Chip
Volume16
Issue24
Pagination4742-4748
Date Published2016 Nov 29
ISSN1473-0189
KeywordsAnimals, Cell Line, Tumor, Cell Separation, Mice, NIH 3T3 Cells, Sequence Analysis, RNA, Single-Cell Analysis
Abstract

Single-cell transcriptome sequencing highly requires a convenient and reliable method to rapidly isolate a live cell into a specific container such as a PCR tube. Here, we report a modular single-cell pipette (mSCP) consisting of three modular components, a SCP-Tip, an air-displacement pipette (ADP), and ADP-Tips, that can be easily assembled, disassembled, and reassembled. By assembling the SCP-Tip containing a hydrodynamic trap, the mSCP can isolate single cells from 5-10 cells per μL of cell suspension. The mSCP is compatible with microscopic identification of captured single cells to finally achieve 100% single-cell isolation efficiency. The isolated live single cells are in submicroliter volumes and well suitable for single-cell PCR analysis and RNA-sequencing. The mSCP possesses merits of convenience, rapidness, and high efficiency, making it a powerful tool to isolate single cells for transcriptome analysis.

DOI10.1039/c6lc01241h
Alternate JournalLab Chip
PubMed ID27841430
PubMed Central IDPMC5554467
Grant ListR01 CA180083 / CA / NCI NIH HHS / United States
R01 DA035868 / DA / NIDA NIH HHS / United States
R21 CA191179 / CA / NCI NIH HHS / United States
R56 AG049714 / AG / NIA NIH HHS / United States

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