Something had stained my cells' nuclei unexpectedly and it can be DAPI. But I wonder how did DAPI stain mix to my samples. Can it mix from my stock solutions which we store at a room temperature? Or somehow from my pipettes? Or should i think on the stains that we store at low temperature?
Hi there, it is easy to check with fluorescence microscopy is it DAPI or not. Probably it came with pipettes (sometimes happens if someone is in rush or not careful) into samples or it contaminated solution(s), which is(are) used for sample preparation. DAPI`s efficiency does not reduced, if its stored at room temperature.
Thank you for your help. Another question of me is if it came with pipettes, can autoclaving our pipettes solve our DAPI problem? (Our pipettes are autoclavable)
I believe that complete cleaning of pipettes with 70% Ethanol solution in destilled water will be enough. To be completly sure you can autoclave them.
I forgot to ask one thing actually. In your original protocol you use DAPI for some reason or not? If you use it and observe your samples at the same day they were prepared, and there is no DAPI response but on the next day it is, then problem is not in pipettes. This phenomenon is OK and it is hard to erradicate completly because some amount of DAPI always binds to the cell membrane and from there due to chemical and physical laws it will get to the nucleus and will stain it. Low temperature slows this effect but it will still occure over some time. Hope this will help.
We do not use DAPI in our lab. However we used to use. So maybe the problem is our stains which (maybe)contain low amounts of DAPI.
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