Dear Researcher, After establishment of the migration Assay based on physical cell exclusion, I was seeing a undesired accumulation of cells in the middle of the cell free gap from up to down (See picture 1: Gap is only shown partly). Cells are already detaching before the incubation actually started. (See picture 2, untreated cells at time point 0h, when I just removed the physicalbarrier between the cell layers).
To understand my procedure before treatment: After removing the barrier, I am washing the cells properly before treating with compounds and due to different imaging setting, they are incubated around 5-10 min in PBS before further treatment.
Can you help me with any idea, how I could reduce these cell accumulations?
Sure, cell accumulation may be increased when migration is also induced, and so cell clusters may be possible. But I am wondering,
1) why my cells detach that early (at time point 0h)
2) Why cells are accumulating in the middle
I am thankful for every input and shared experiences!
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