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Drop-on-Demand Single Cell Isolation and Total RNA Analysis

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  • Sangjun Moon
  • Yun-Gon Kim
  • Lingsheng Dong
  • Michael Lombardi
  • Edward Haeggstrom
  • Roderick V Jensen
  • Li-Li Hsiao
  • Utkan Demirci

Abstract

Technologies that rapidly isolate viable single cells from heterogeneous solutions have significantly contributed to the field of medical genomics. Challenges remain both to enable efficient extraction, isolation and patterning of single cells from heterogeneous solutions as well as to keep them alive during the process due to a limited degree of control over single cell manipulation. Here, we present a microdroplet based method to isolate and pattern single cells from heterogeneous cell suspensions (10% target cell mixture), preserve viability of the extracted cells (97.0±0.8%), and obtain genomic information from isolated cells compared to the non-patterned controls. The cell encapsulation process is both experimentally and theoretically analyzed. Using the isolated cells, we identified 11 stem cell markers among 1000 genes and compare to the controls. This automated platform enabling high-throughput cell manipulation for subsequent genomic analysis employs fewer handling steps compared to existing methods.

Suggested Citation

  • Sangjun Moon & Yun-Gon Kim & Lingsheng Dong & Michael Lombardi & Edward Haeggstrom & Roderick V Jensen & Li-Li Hsiao & Utkan Demirci, 2011. "Drop-on-Demand Single Cell Isolation and Total RNA Analysis," PLOS ONE, Public Library of Science, vol. 6(3), pages 1-10, March.
    • Handle: RePEc:plo:pone00:0017455
    DOI: 10.1371/journal.pone.0017455
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    References listed on IDEAS

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    1. Paolo Bianco & Pamela Gehron Robey, 2001. "Stem cells in tissue engineering," Nature, Nature, vol. 414(6859), pages 118-121, November.
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    Cited by:

    1. SangJun Moon & Elvan Ceyhan & Umut Atakan Gurkan & Utkan Demirci, 2011. "Statistical Modeling of Single Target Cell Encapsulation," PLOS ONE, Public Library of Science, vol. 6(7), pages 1-11, July.

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