Fluorescent “barcodes” created by labelling pools of cells with different combinations of dyes could be a boon to researchers interested in performing large-scale cell-based studies for drug discovery and other applications, according to Gary Nolan and colleagues at Stanford.
Protein phosphorylation has a major role in a wide variety of essential cellular functions, and several years ago Nolan and colleagues developed “phospho flow”, an approach for simultaneously characterizing the phosphorylation status of multiple proteins in different groups of cells. However, phospho flow can become impractical when scaled up for the analysis of multiple protein targets in response to a large variety of different compounds.
Now, Nolan and Peter Krutzik have found a new solution to this problem. They have demonstrated that cells labelled with different concentrations of a fluorophore can be readily distinguished, and that combinations of fluorophores – each present at a different concentration – can generate cell-specific “barcodes”. Thanks to these barcodes, researchers can subject large numbers of cell populations to different treatments, then pool the cells for simultaneous phospho flow sorting and analysis, resulting in a considerable savings in efficiency and reagent consumption.
The team shows that with just three barcoding fluorophores, one can easily sort out the phospho flow data for differentially treated cells from a 96-well assay plate – and larger assays should prove equally feasible.
More information in the journal Nature Methods. You can get a free subscription to the print edition of Nature Methods here.