Hi! Recently we started using Cytation 1 in our lab to count cells and see them on a plate. I have no problems counting HEK 293T cells in 24 well plates from "Greiner bio-one" or "Thermo Nunclon Delta Surface" (look at them, they look so pretty!). But when we move on in a 48 well plate from the same brands, Cytation 1 cannot focus, and we have results like the ones in the second picture.
Also, my colleagues are having problems with the 24 well plate but counting fibroblast instead of HEK. We didn't change a thing in the settings, and we are really mad about it. Any ideas?
Thanks!
This seems an autofocus problem when plate was changed. Change back to 24 well plate and see you get sharp image.
1. If yes, then, check user manual for Autofocus methods after putting 48 well plate onto reader.
2. If No, then, check the plate in another reader if you have, or restart the software.
In any case, the Autofocus needs to be calibrated for different plates, as the dimensions, depth etc of these plates differ. Alternatively, the software might have user controlled manual focus settings to fix it at certain height to scan for different plates.
Dear Ana, please can you manually adjust focus and exposure? Looking at the second image, it is evident that you are just two or three clicks away from clarity. Autofocusing might not give same clarity in a 24 well plate as in the 48 well plate since the focusing distances are bound to change. Manually focusing and exposing will do the trick.
Every cell will have its own morphological properties. Hence you can't use the same parameters for different types of cells. Image 2 suggest that the cells are out of focus, hence might be looking blurred. Better play with the instrument parameters.
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