We want to analyse cell migration by using the ClearView Chemotaxis Assay from Incucyte. We put the cells to incubate for 24 hours and after each hour a photo was taken.
At timepoint 0 the Incucyte focusses correctly on the membrane --> photo1
After a couple of hours it doesn't anymore --> photo 2+3.
And this doesn't just happen to one well, it happens to most/all of them.
Sometimes one hour later it focusses correctly again. This happened overnight, so it's very unlikely someone accidentally moved my plate.
Does anyone know why it sometimes doesn't focus corretcly or how it can be resolved?
It seemed that quite a lot of the medium in the reservoir was evaporated, eventhough we used the recommended 200ul. So maybe a bubble somehow formed causing the membrane to be dry?
Any other ideas?
Thanks a lot!