Digital microfluidic isolation of single cells for -Omics

被引:122
作者
Lamanna, Julian [1 ,2 ]
Scott, Erica Y. [1 ,2 ]
Edwards, Harrison S. [2 ,3 ]
Chamberlain, M. Dean [1 ,2 ]
Dryden, Michael D. M. [1 ]
Peng, Jiaxi [1 ]
Mair, Barbara [2 ]
Lee, Adam [3 ]
Chan, Calvin [1 ]
Sklavounos, Alexandros A. [1 ,2 ]
Heffernan, Austin [2 ]
Abbas, Farhana [1 ,2 ]
Lam, Charis [1 ]
Olson, Maxwell E. [1 ]
Moffat, Jason [2 ,3 ,4 ]
Wheeler, Aaron R. [1 ,2 ,3 ]
机构
[1] Univ Toronto, Dept Chem, 80 St George St, Toronto, ON M5S 3H6, Canada
[2] Univ Toronto, Donnelly Ctr Cellular & Biomol Res, 160 Coll St, Toronto, ON M5S 3E1, Canada
[3] Univ Toronto, Inst Biomat & Biomed Engn, 164 Coll St, Toronto, ON M5S 3G9, Canada
[4] Univ Toronto, Dept Mol Genet, Toronto, ON M5S 3H6, Canada
基金
加拿大自然科学与工程研究理事会;
关键词
GENOME-WIDE EXPRESSION; GENE-EXPRESSION; RNA-SEQ; PLATFORM; AMPLIFICATION; SUCCESS;
D O I
10.1038/s41467-020-19394-5
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
We introduce Digital microfluidic Isolation of Single Cells for -Omics (DISCO), a platform that allows users to select particular cells of interest from a limited initial sample size and connects single-cell sequencing data to their immunofluorescence-based phenotypes. Specifically, DISCO combines digital microfluidics, laser cell lysis, and artificial intelligence-driven image processing to collect the contents of single cells from heterogeneous populations, followed by analysis of single-cell genomes and transcriptomes by next-generation sequencing, and proteomes by nanoflow liquid chromatography and tandem mass spectrometry. The results described herein confirm the utility of DISCO for sequencing at levels that are equivalent to or enhanced relative to the state of the art, capable of identifying features at the level of single nucleotide variations. The unique levels of selectivity, context, and accountability of DISCO suggest potential utility for deep analysis of any rare cell population with contextual dependencies. Multi-Omic approaches are a powerful way for obtaining in-depth understanding of a cell's state. Here the authors present DISCO, combining digital microfluidics, laser cell lysis, and artificial intelligence-driven image processing to analyze single-cell genomes, transcriptomes and proteomes in a mixed population.
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页数:13
相关论文
共 68 条
[1]   Gene essentiality and synthetic lethality in haploid human cells [J].
Blomen, Vincent A. ;
Majek, Peter ;
Jae, Lucas T. ;
Bigenzahn, Johannes W. ;
Nieuwenhuis, Joppe ;
Staring, Jacqueline ;
Sacco, Roberto ;
van Diemen, Ferdy R. ;
Olk, Nadine ;
Stukalov, Alexey ;
Marceau, Caleb ;
Janssen, Hans ;
Carette, Jan E. ;
Bennett, Keiryn L. ;
Colinge, Jacques ;
Superti-Furga, Giulio ;
Brummelkamp, Thijn R. .
SCIENCE, 2015, 350 (6264) :1092-1096
[2]   Comparison of whole genome amplification techniques for human single cell exome sequencing [J].
Borgstrom, Erik ;
Paterlini, Marta ;
Mold, Jeff E. ;
Frisen, Jonas ;
Lundeberg, Joakim .
PLOS ONE, 2017, 12 (02)
[3]   Intelligent image-based in situ single-cell isolation [J].
Brasko, Csilla ;
Smith, Kevin ;
Molnar, Csaba ;
Farago, Nora ;
Hegedus, Lili ;
Balind, Arpad ;
Balassa, Tamas ;
Szkalisity, Abel ;
Sukosd, Farkas ;
Kocsis, Katalin ;
Balint, Balazs ;
Paavolainen, Lassi ;
Enyedi, Marton Z. ;
Nagy, Istvan ;
Puskas, Laszlo G. ;
Haracska, Lajos ;
Tamas, Gabor ;
Horvath, Peter .
NATURE COMMUNICATIONS, 2018, 9
[4]   Clarifying intact 3D tissues on a microfluidic chip for high-throughput structural analysis [J].
Chen, Yih Yang ;
Silva, Pamuditha N. ;
Syed, Abdullah Muhammed ;
Sindhwani, Shrey ;
Rocheleau, Jonathan V. ;
Chan, Warren C. W. .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2016, 113 (52) :14915-14920
[5]   Hydro-Seq enables contamination-free high-throughput single-cell RNA-sequencing for circulating tumor cells [J].
Cheng, Yu-Heng ;
Chen, Yu-Chih ;
Lin, Eric ;
Brien, Riley ;
Jung, Seungwon ;
Chen, Yu-Ting ;
Lee, Woncheol ;
Hao, Zhijian ;
Sahoo, Saswat ;
Kang, Hyun Min ;
Cong, Jason ;
Burness, Monika ;
Nagrath, Sunitha ;
Wicha, Max S. ;
Yoon, Euisik .
NATURE COMMUNICATIONS, 2019, 10 (1)
[6]   Digital Microfluidics [J].
Choi, Kihwan ;
Ng, Alphonsus H. C. ;
Fobel, Ryan ;
Wheeler, Aaron R. .
ANNUAL REVIEW OF ANALYTICAL CHEMISTRY, VOL 5, 2012, 5 :413-440
[7]   In Silico Labeling: Predicting Fluorescent Labels in Unlabeled Images [J].
Christiansen, Eric M. ;
Yang, Samuel J. ;
Ando, D. Michael ;
Javaherian, Ashkan ;
Skibinski, Gaia ;
Lipnick, Scott ;
Mount, Elliot ;
O'Neil, Alison ;
Shah, Kevan ;
Lee, Alicia K. ;
Goyal, Piyush ;
Fedus, William ;
Poplin, Ryan ;
Esteva, Andre ;
Berndl, Marc ;
Rubin, Lee L. ;
Nelson, Philip ;
Finkbeiner, Steven .
CELL, 2018, 173 (03) :792-+
[8]   U87MG Decoded: The Genomic Sequence of a Cytogenetically Aberrant Human Cancer Cell Line [J].
Clark, Michael James ;
Homer, Nils ;
O'Connor, Brian D. ;
Chen, Zugen ;
Eskin, Ascia ;
Lee, Hane ;
Merriman, Barry ;
Nelson, Stanley F. .
PLOS GENETICS, 2010, 6 (01)
[9]   Improved Single-Cell Proteome Coverage Using Narrow-Bore Packed NanoLC Columns and Ultrasensitive Mass Spectrometry [J].
Cong, Yongzheng ;
Liang, Yiran ;
Motamedchaboki, Khatereh ;
Huguet, Romain ;
Thy Truong ;
Zhao, Rui ;
Shen, Yufeng ;
Lopez-Ferrer, Daniel ;
Zhu, Ying ;
Kelly, Ryan T. .
ANALYTICAL CHEMISTRY, 2020, 92 (03) :2665-2671
[10]  
DeCarlo K, 2011, METHODS MOL BIOL, V755, P1, DOI 10.1007/978-1-61779-163-5_1