Abstract
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.
Highlights
Characterization of biological samples relies heavily on microscopy where, in response to various stimuli, molecular probes and a myriad of contrast reagents are routinely used to identify and label individual live cells of interest
To the best of our knowledge, single cell magneto-optical capture (scMOCa) is the only technology that permits isolation, and subsequent clonal expansion, of extremely small numbers of cells from relatively large heterogeneous populations based solely on visual criteria. scMOCa is highly efficient, as the fraction of tagged cells collected in the top chamber exhibits minimal capture losses and high specificity
Adaptations needed for capturing cell populations representing
Summary
Characterization of biological samples relies heavily on microscopy where, in response to various stimuli, molecular probes and a myriad of contrast reagents are routinely used to identify and label individual live cells of interest These methods often require prior knowledge of cellular markers or use of elaborate reporter constructs. We recently developed a method termed Cell Labeling via Photobleaching (CLaP) (Binan et al, 2016) allowing the arbitrary tagging of individual cells among a heterogeneous population within a microscopy field This is accomplished by crosslinking biotin molecules to their plasma membranes with the lasers of a confocal microscope, followed by use of fluorescent streptavidin conjugates to reveal the marked cells. The ease of use and affordability of our method is expected to facilitate the characterization of phenotypes of interest occurring in a small fraction of cell populations
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