Hi all,
I recently started working with flow cytometry with no prior knowledge. I have been trying to stain cells to practice flow cytometry techniques, such as panel setup, gating, and data analysis.
I have come to realize that the cells I use for practice don't particularly express the proteins that my marker antibodies selectively bind to. The sample prep itself is also tedious and not super efficient considering that I want to learn about the practical part of the instrument.
I want to use fluorescent beads for practice but am swamped with endless choices. Could you recommend two types of beads that are different in size and fluorescence labeling? If you have budget-friendly options, that'd be even better.
In case you know of other ways to easily practice flow cytometry operations, I'd be glad to hear them. Thanks in advance for your help.
Hi Wey,
Although I am not an expert in Flow Cytometry I can tell you that some F.C. collegues and myself use the reagents from BD Bioscience: https://www.bdbiosciences.com/en-us/products/reagents/flow-cytometry-reagents
Good efficiency, low autofluorescence and also pretty trustworthy to gate.
Regarding the specific topic you comment of the fluorescence beads... maybe you can consider using classic FITC or PE conjugated antibodies to gain expertise (2 fluorophors with very well reported activity and data).
I hope this helps!
Francesc Estrany
Hi Wey,
A great way to practice with flow is to use simple stains like Annexin V and PI (measures of early and late apoptosis). The staining protocol is only about 30 minutes as you don't wash out the stain. You can purposely spike in dead cells to make sure you have positive and negative populations. Another suggestion would be to mix two cell lines with different size/morphology so you can practice gating with FSC/SSC.
For beads BD makes great reagents all the way around. I also like ThermoFishers Ultra comp beads (01-3333-41) which would allow you to use the same colors you'll be using in the future.
Best of luck!
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