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Optical painting and fluorescence activated sorting of single adherent cells labelled with photoswitchable Pdots

Author

Listed:
  • Chun-Ting Kuo

    (University of Washington)

  • Alison M. Thompson

    (University of Washington)

  • Maria Elena Gallina

    (University of Washington)

  • Fangmao Ye

    (University of Washington)

  • Eleanor S. Johnson

    (University of Washington)

  • Wei Sun

    (University of Washington)

  • Mengxia Zhao

    (University of Washington)

  • Jiangbo Yu

    (University of Washington)

  • I-Che Wu

    (University of Washington)

  • Bryant Fujimoto

    (University of Washington)

  • Christopher C. DuFort

    (Fred Hutchinson Cancer Research Center)

  • Markus A. Carlson

    (Fred Hutchinson Cancer Research Center)

  • Sunil R. Hingorani

    (Fred Hutchinson Cancer Research Center)

  • Amy L. Paguirigan

    (Fred Hutchinson Cancer Research Center)

  • Jerald P. Radich

    (Fred Hutchinson Cancer Research Center)

  • Daniel T. Chiu

    (University of Washington)

Abstract

The efficient selection and isolation of individual cells of interest from a mixed population is desired in many biomedical and clinical applications. Here we show the concept of using photoswitchable semiconducting polymer dots (Pdots) as an optical ‘painting’ tool, which enables the selection of certain adherent cells based on their fluorescence, and their spatial and morphological features, under a microscope. We first develop a Pdot that can switch between the bright (ON) and dark (OFF) states reversibly with a 150-fold contrast ratio on irradiation with ultraviolet or red light. With a focused 633-nm laser beam that acts as a ‘paintbrush’ and the photoswitchable Pdots as the ‘paint’, we select and ‘paint’ individual Pdot-labelled adherent cells by turning on their fluorescence, then proceed to sort and recover the optically marked cells (with 90% recovery and near 100% purity), followed by genetic analysis.

Suggested Citation

  • Chun-Ting Kuo & Alison M. Thompson & Maria Elena Gallina & Fangmao Ye & Eleanor S. Johnson & Wei Sun & Mengxia Zhao & Jiangbo Yu & I-Che Wu & Bryant Fujimoto & Christopher C. DuFort & Markus A. Carlso, 2016. "Optical painting and fluorescence activated sorting of single adherent cells labelled with photoswitchable Pdots," Nature Communications, Nature, vol. 7(1), pages 1-11, September.
    • Handle: RePEc:nat:natcom:v:7:y:2016:i:1:d:10.1038_ncomms11468
    DOI: 10.1038/ncomms11468
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