Do you have to use such a large vessel? If you interested purely in comparing adhesiveness of different substrates your cells don't have to be fully confluent. You can seed fixed number of labelled cells (10-30 thousands) on coverslips, give them time to adhere, wash off unbound cells and count whatever is left under the microscope. It should answer your question and shouldn't take too much time. Hope that helps

Sylwia

Can you label your cells and then score them using a microscope? that should give you precise number of adherent cells. CellTracker™ dyes are quick to label, non-toxic and suitable for live cell experiments. Alternatively, if you cells don't have to be alive, you can fix them with formalin and do immunostaining

Thanks for the comment Kabacik. That's actually not a bad idea. Although I'll be skeptical about how precise the counting can be since my substrate is 3cm x 3cm and at fully confluence stage, they will be a lot of cells, making the process tedious and inaccurate.

Do you have to use such a large vessel? If you interested purely in comparing adhesiveness of different substrates your cells don't have to be fully confluent. You can seed fixed number of labelled cells (10-30 thousands) on coverslips, give them time to adhere, wash off unbound cells and count whatever is left under the microscope. It should answer your question and shouldn't take too much time. Hope that helps

Sylwia

Hello Udesh,

I agree with Sylwia answer above and would say that if you are interested to evaluate cell adhesion you won´t need to cultivate your cells for days. I think you should perform your incubation for 60-90 minutes, wash non-adherent cells and then proceed with counting. Regarding your concern about cell viability, in this case you should use MTT to quantify your adhesion assay.

Hope it helps.

I agree with Sylwia and Newton. I suggest to consider the time required for cell adhesion and proliferation as it may vary with cell lines.

Thanks for your comments, Slywia, Newton and Syed. The problem remains: How to accurately count labelled adhered cells?

as I said before, if you score the cells under microscope, the number of cells per field of view will give you accurate number of cells adhering to your substrate

Thanks Sylvia. I'll give it a try.