In addition to an equal cell number, a homogeneous cell distribution on the growth surface of a culture vessel can have an impact on your experimental outcome. Different techniques are common, usually passed from more experienced scientists in a lab to their trainees. But what is the best method to evenly distribute the cells on the growth surface? In Image 3, you can see a comparison of different techniques.
On the left, you see what happens if you just prefill the vessel with cell culture medium, add the cells, and call it a day. As a result, the cells mainly adhere in the middle of the vessel. To achieve an equal cell distribution, some people use a “figure-eight” movement, while others prefer a cross-like movement of the plate or dish. Both techniques lead to a better distribution of the cells. However, the smaller the vessel diameter, the less movement of the liquid you have, leading to an uneven distribution of cells. An effective way to circumvent this effect during cell seeding is to first dilute the cell suspension to the desired concentration in a tube or reservoir (mastermix) and then pipette the final volume to the culture vessel in one step.
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