Optical painting and fluorescence activated sorting of single adherent cells labelled with photoswitchable Pdots.
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.
Department of Chemistry, University of Washington, Seattle, Washington 98195, USA.
Chun-Ting
CT
Kuo
Department of Chemistry, University of Washington, Seattle, Washington 98195, USA.
Alison M
AM
Thompson
Department of Chemistry, University of Washington, Seattle, Washington 98195, USA.
Maria Elena
ME
Gallina
Department of Chemistry, University of Washington, Seattle, Washington 98195, USA.
Fangmao
F
Ye
Department of Chemistry, University of Washington, Seattle, Washington 98195, USA.
Eleanor S
ES
Johnson
Department of Chemistry, University of Washington, Seattle, Washington 98195, USA.
Wei
W
Sun
Department of Chemistry, University of Washington, Seattle, Washington 98195, USA.
Mengxia
M
Zhao
Department of Chemistry, University of Washington, Seattle, Washington 98195, USA.
Jiangbo
http://orcid.org/0000-0001-6708-9781
J
Yu
Department of Chemistry, University of Washington, Seattle, Washington 98195, USA.
I-Che
IC
Wu
Department of Chemistry, University of Washington, Seattle, Washington 98195, USA.
Bryant
B
Fujimoto
Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA.
Christopher C
CC
DuFort
Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA.
Markus A
MA
Carlson
Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA.
Sunil R
SR
Hingorani
Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA.
Amy L
AL
Paguirigan
Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA.
Jerald P
JP
Radich
Department of Chemistry, University of Washington, Seattle, Washington 98195, USA.
Daniel T
DT
Chiu
D005453:Fluorescence; D006801:Humans; D061986:MCF-7 Cells; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D057054:Molecular Imaging / Q000379:methods; D011108:Polymers / Q000737:chemistry*; D045663:Quantum Dots; D012666:Semiconductors
D011108:Polymers