Currently, I am trying to see the chemotaxis of naive T cells on ICAM-1 coated plates. Although they adhere well after few minutes of CCL19 treatment, I mostly see the flow of the cells without adhesion during the very first minutes. How can I get rid of this movement?

Here is how I do my exp;

- Coat glass-bottom confocal dishes with 10ug/ml ICAM-1 Fc overnight.

- Next day, I seed 1x10^5 cells and spin them for 1 min to adhere them more (500rpmi) Then, incubate in an incubator for 5 min to stabilize cells

- I use EVOS fl auto 2 imaging system with an incubator (37C and 5%CO2)

- I place cells and arrange the plate for imaging. After choosing the right area, I treat cells 10ug CCL19 and image for 15 minutes.

From the video, you can see that cells initially flow from northeast to southwest. This video clip is from the first 5 min of treatment of CCL19.