Fluorescent cell barcoding in flow cytometry

by Alexey Bersenev on November 13, 2011 · 0 comments

in cytometry, methods

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Fluorescent cell barcoding (FBC) was developed in Nolan’s lab at Stanford University a few years ago. For the first time, they used it for studying signaling pathways in activated/ inhibited T-cells. This technique is very promising and could find a wide applicability in stem cell research.

Definition and advantages:

Fluorescent cell barcoding (FCB) enables high throughput, high content flow cytometry by multiplexing samples prior to staining and acquisition on the cytometer. Individual cell samples are barcoded, or labeled, with unique signatures of fluorescent dyes so that they can be mixed together, stained, and analyzed as a single sample. By mixing samples prior to staining, antibody consumption is typically reduced 10- to 100-fold. In addition, data robustness is increased through the combination of control and treated samples, which minimizes pipetting error, staining variation, and the need for normalization. Finally, speed of acquisition is enhanced, enabling large profiling experiments to be run with standard cytometer hardware.

Typical FCB phospho-flow scheme and panel you can look at here.

FBC method was picked up by other labs and optimized. Recently, FBC was used for studying of cytokine signaling in T-cells from small volume of patient’s blood. The authors optimized FBC method in order to avoid some technical considerations. I think, fluorescent cell barcoding should be widely used to study signaling pathways in stem cell research.

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