Good day to everyone. I ask your help to solve some questions about the FACS
canto II cytometer. It turns out that when starting
a new experiment after some time the compensations and
the parameters are erased or disabled. some of the most
common are APC, Pacific Blue and Percp or Percep cy5.5
are disabled and the SSC and FSC change the value of
the established parameter. Someone suggested that it
could be the configuration but it has not moved,
I have no idea what it can be or what causes these
errors to appear. When I create a new experiment and stable the
fluorochromes with which it is going to work,
it also happens that I start to use other fluorochromes that I do not need, for example, I delimit six fluorochromes APC, FITC, PB, PE, Percp, APCcy7 ... a sometimes also comes Percpcy5.5, Alexa fluor 647 ... etc. The engineer has already checked the cytometer but I still have these conflicts Do I do something wrong? Can someone tell me why this is due? How can I solve that? I leave some representative images Thank you.
Have you checked if you have multiple configurations? perhaps it is changing from one to another, or you are selecting the wrong one when doing the CS&T. Also, I suggest adjusting the "parameters (channels)" in "cytometry settings" in the default tube that comes out when you add a new "specimen", adjust the autofluorescence in that tube, and then copy it and paste it as many tubes as you need. If you need more help, feel free to message me, good luck.
As Lucinda indicated, it is very probable that you have several configurations. This is common when there are several users from distinct labs. Besides checking the configuration, you also should check if the filters for your missing fluorochromes are installed in the FACSCanto II.
In my experience, if the configuration is all right, you can open a previous experiment with your desired configuration; then duplicate this experiment (either with the right click of the mouse or by clicking Edit in the upper tab of the Diva). The duplicated experiment will contain your configuration, PMT values, compensation tubes and specimens. If you cannot duplicate it, it will appear a message indicating a problem with the configuration of your cytometer.
Hope this helps and good luck
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