Hi dear friends,
I am working on Parkinson model. I transfected SH-SY5Y cells with topo isomerase IIB by lipofectamine 3000 reagent. I took some photos today but their backgrounds were too noisy. Notably, I took them by different channels, but I have got the same result. Worth to mention that we are sing ZEISS.
My question is what can I do to solve this problem?
I have added photos below.
Thanks in advance.
Hi,
I think this is a typical photograph.
I think the higher noise is due to the lower fluorescence intensity and longer exposure time.
Therefore, the magnification of the lens should be set to a higher magnification. Or, it would be better to use a confocal microscope.
Or, although it is not a fundamental solution, you can change the contrast according to the appropriate standard.
Hello,
first I want to ask a question, why you have a very low number of cells? the noise can come from a lack of washing before and/or after the fixation step or the mounting step...also, did you clean properly your slides? Maybe it is seems less probable, but you have to also check the cleaness of your microscope's lense(s).
Narumi Uno Thank you for your suggestions.
Iskander Madhi I have not stained them yet, it is GFP after transfection. Thank you for your recommendations it would of help in the next step of my experiment.
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