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

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. Carlson, Sunil R. Hingorani, Amy L. Paguirigan, Jerald P. Radich and Daniel T. Chiu ()
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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

Nature Communications, 2016, vol. 7, issue 1, 1-11

Abstract: 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.

Date: 2016
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DOI: 10.1038/ncomms11468

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