cancer biology; cell biology; cell labeling; cell sorting; clonal expansion; none; rare cells; single cell capture; visual phenotypes; Streptavidin; Animals; Cell Line; Cell Separation/methods; Humans; Staining and Labeling/methods; Streptavidin/metabolism; Magnetic Fields; Cell Separation; Staining and Labeling; Neuroscience (all); Biochemistry, Genetics and Molecular Biology (all); Immunology and Microbiology (all)
Abstract :
[en] The ability to isolate rare live cells within a heterogeneous population based solely on visual criteria remains technically challenging, due largely to limitations imposed by existing sorting technologies. Here, we present a new method that permits labeling cells of interest by attaching streptavidin-coated magnetic beads to their membranes using the lasers of a confocal microscope. A simple magnet allows highly specific isolation of the labeled cells, which then remain viable and proliferate normally. As proof of principle, we tagged, isolated, and expanded individual cells based on three biologically relevant visual characteristics: i) presence of multiple nuclei, ii) accumulation of lipid vesicles, and iii) ability to resolve ionizing radiation-induced DNA damage foci. Our method constitutes a rapid, efficient, and cost-effective approach for isolation and subsequent characterization of rare cells based on observable traits such as movement, shape, or location, which in turn can generate novel mechanistic insights into important biological processes.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Binan, Loïc; Research Center, Maisonneuve-Rosemont Hospital, Montreal, Canada ; Department of Ophthalmology, University of Montreal, Montreal, Canada
Bélanger, François; Research Center, Maisonneuve-Rosemont Hospital, Montreal, Canada ; Department of Medicine and Molecular Biology Program, University of Montreal, Montreal, Canada
Uriarte, Maxime ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques ; Research Center, Maisonneuve-Rosemont Hospital, Montreal, Canada ; Department of Medicine and Molecular Biology Program, University of Montreal, Montreal, Canada
Lemay, Jean François ; Research Center, Maisonneuve-Rosemont Hospital, Montreal, Canada
Pelletier De Koninck, Jean Christophe; Research Center, Maisonneuve-Rosemont Hospital, Montreal, Canada
Roy, Joannie; Research Center, Maisonneuve-Rosemont Hospital, Montreal, Canada
Affar, El Bachir; Research Center, Maisonneuve-Rosemont Hospital, Montreal, Canada ; Department of Medicine and Molecular Biology Program, University of Montreal, Montreal, Canada
Drobetsky, Elliot; Research Center, Maisonneuve-Rosemont Hospital, Montreal, Canada ; Department of Medicine and Molecular Biology Program, University of Montreal, Montreal, Canada
Wurtele, Hugo; Research Center, Maisonneuve-Rosemont Hospital, Montreal, Canada
Costantino, Santiago ; Research Center, Maisonneuve-Rosemont Hospital, Montreal, Canada ; Department of Ophthalmology, University of Montreal, Montreal, Canada
Language :
English
Title :
Opto-magnetic capture of individual cells based on visual phenotypes.
CIHR - Canadian Institutes of Health Research NSERC - Natural Sciences and Engineering Research Council FRQS - Fonds de Recherche du Québec - Santé Canadian Cancer Society FRQNT - Fonds de Recherche du Québec - Nature et Technologies
Funding text :
We thank Maryam Tabatabaei and Sébastien Talbot (Université de Montréal) for help with experiments using DRG cells. This work was supported by grants from the Natural Science and Engineering Research Council of Canada to ED, EBA, SC and HW, Genome Canada/Génome Québec and Canadian Cancer Society to SC, Fonds de Recherche du Québec – Nature et Technologies to SC, and the Canadian Institutes of Health Research to ED, EBA and HW SC, EBA and HW hold salary awards from the Fonds de Recherche du Québec – Santé.Canadian Institutes of Health Research to El Bachir Affar, Elliot Drobetsky, Hugo Wurtele. Natural Sciences and Engineering Research Council of Canada to El Bachir Affar, Elliot Drobetsky, Hugo Wurtele, Santiago Costantino. Fonds de Recherche du Qu?bec - Sant? to El Bachir Affar, Hugo Wurtele. Genome Canada to Santiago Costantino. Canadian Cancer Society to Santiago Costantino. Fonds de Recherche du Qu?bec - Nature et Technologies to Santiago Costantino.
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