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Our Scientific Support team share some additional flow cytometry troubleshooting tips from a recent workshop.

In order to better serve our customers, the members of the Abcam scientific support team frequently attend training sessions that increase and refresh our technical knowledge. In October, Elena, Jackie, and Caitlin from the US team completed a day-long flow cytometry workshop at the Flow Cytometry Core of Beth Israel Deaconess Medical Center in Boston, under the instruction of technical director John Tigges. The day started with a brief history of fluorescence and flow cytometry (the first cytometer was invented in 1968!) and continued with in-depth training on the fluidics, optics, and electronics systems involved in the machines. John was kind enough to also give us a full afternoon running fluorescent beads on an LSRII!

What we learned

• When immunostaining for proteins that will be rare in the cell population, use a stronger fluorophore like phycoerythrin (PE).
• Don't change any voltages once they're set to the unstained control.
• If the sample pressure is too high, multiple cells will be analyzed at once. This is fine for IF analysis, but if the CV's are important (such as for cell cycle or ploidy studies) then this needs to be corrected.
• Prevent air from being introduced into the system, otherwise the pressure won't be sufficient and events won't be seen.

» For more tips, visit our previous blog article: Top 10 tips for flow cytometry