In my lab, we are only a few ppl running multicolor panels in a 14 color BD Fortessa. Most users only use 1 color and therefor does not run CST beads. My question is if it's recommended to run the CST beads on a regular basis, or if it's ok to only run them before the acquisition of a multicolor panel?
Cheers,
Annette
Annette, CST does not 'improve' the cytometer's performance. But it does track the instruments performance over time. It is highly recommended to run CST each time the instrument is turned on or the instrument configuration file is changed. CST provides data regarding laser delay values, stability of signal detection for each laser and detector in terms of signal intensity as well as resolution of dim signals from the negative. It also calculates the optimal detector voltages based on that day's performance which can then be used to define an application settings file for your experiment, ensuring that changes in signal intensity are a result of the experiment and not the instrument! Imagine this scenario - you are running a single color experiment over time and the signal tends to be dim to moderately bright and then one day the positive signal cannot be distinguished from the negative signal. Are any of these differences real or could they be a product of a failing detector or laser or a dirty flow cell? How do you know if you are not consistently monitoring performance? I know it can be a pain to wait for the CST program to open, but running the performance check only takes about 10min. Isn't your data valuable enough to take that extra time? Don't you want to be able to foresee potential instrument problems and address them BEFORE a complete failure? Think about it....
Thanks for the answer Lynda! I always run the CST, but the ppl running only one color dont... We are thinking of making it mandatory to run CST, but if running it 1-2 times a week is enough, the argument of enhanced performance is not valid. I'm interested in having a perfectly functional machine to run my samples in and my question really is about if the stability of the lasers are enhanced by frequently running CST or if it doesnt matter. What the others do with their samples is up to them...
As Lynda said it is highly recommended to run CSTs every day you use your flow cytometry even if it is for analyzing one color. The stability of the lasers is not affected by running the CSTs, but you can have data of how is the performance of your instrument, which can be visualized in Levey-Jennings plots that provides the software. With these data you can evaluate if the lasers and every PMT detector (including those that you or your colleagues do not use) are correctly working. Thus, in case of any malfunctioning, it is easier to make a diagnostic with this information.
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