I'm new to Flow cytometry and was wondering if there is a way to find out which are your fluorophores, when the samples are not labeled, just looking to the graphs. I had some problem with the FACScalibur and my samples were not stored with names, just with sequential numbers. Then unless I have how to identify my fluorophores, I'll lose this experiment.
Does anyone have an input about that? Thanks
Best way is to use compensation beads available with BD Biosciences possibly by others. Choose the right one which depends on the host in which the antibodies of interest generated. This is pretty standard way that people follow especially when the supply of the cells of interest is very limited.
I have definitely misunderstood the question, please accept my apologies.
Yes, I agree with Ceri. Another way around is to repeat the experiment n store the data with proper labeling and compare the results from the expt that was not labeled for fluorophores.
Thank you both for your comments. Yes, unfortunately I had trouble with the CellQuest and I don't have the right names on file. If the problem was just that was ok, because I know the sequence I ran the samples. The problem is that I have 3 times the number of samples I ran, so I don't know what happen with the software...
Anyway, since there is no way to figure it out which are the compensation samples (I have 4 fluorophores, FITC, PE, PerCP and APC), I'll discard this experiment and perform it again.
Hey Sabrina,
I'm wondering if this is an issue with FlowJo not having the preferences set so that it shows the Sample ID and Tube ID? If so, then you might try and change the way that FlowJo names your files, it can be a mix of keywords written to your FCS file or short file name.
Let me know if we can help with this or with your data here or at [email protected]
Thanks,
Miguel Velazquez
FlowJo TechSupport
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