Title | Single-cell isolation by a modular single-cell pipette for RNA-sequencing. |
Publication Type | Journal Article |
Year of Publication | 2016 |
Authors | Zhang, K, Gao, M, Chong, Z, Li, Y, Han, X, Chen, R, Qin, L |
Journal | Lab Chip |
Volume | 16 |
Issue | 24 |
Pagination | 4742-4748 |
Date Published | 2016 Nov 29 |
ISSN | 1473-0189 |
Keywords | Animals, 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. |
DOI | 10.1039/c6lc01241h |
Alternate Journal | Lab Chip |
PubMed ID | 27841430 |
PubMed Central ID | PMC5554467 |
Grant List | R01 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 |
Single-cell isolation by a modular single-cell pipette for RNA-sequencing.
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