i would like to investigate the movement of neutrophil in 96 well plate and i need to coat the well.
does anyone have experience about the best coat for neutrophil movement? BSA? HSA? FBS? PDL?
Thanks
There are a few different coatings you can use for this:
I used poly-l-lysine, which worked very well and allowed the neutrophils to attach to the surface and also remain very motile. The drawback of using poly-l-lysine, is that neutrophils would not encounter this in-vivo and so it might not be suited for your experiments.
A more realistic representation of an in-vivo situation would involve using a coating such as gelatine or ICAM.
We measure neutrophil migration across cover slips, and we use BSA as a surrogate, as recommended in this paper:
http://www.sciencedirect.com/science/article/pii/S0022175900001927
IF YOU COAT THE WELLS BY AGAROSE GEL YOU CAN SEE NEUTROPHİL MOVEMENTS UNDER AGAROSE. THE END OF STUDY YOU WİLL GET OUT AGAROSE GEL AND STAIN WELLS TO SEE MOVEMENT TRACES ON THE WELLS.
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