Hacat Seeding into 8-well chamber slide - (Feb/13/2012 )

Hi everyone.

I'm pretty new to cell culture work and am trying to work something out.

I want to seed HaCats into 8-well culture/chamber slides (http://www.emsdiasum.com/microscopy/products/preparation/chamber.aspx)
After 72 hours I want to harvest them for immunofluorescent staining.

I am wondering how many cells I should put into each well so that they are essentially confluent after 72 hours

Any ideas?
I think the area in each well is ~0.8-1cm2

Thankyou for your time

thommo

It depends a bit on how big HaCats are and how fast they grow, but at confluency you will have a little bit less than 10⁵ cells in each well. If you know how fast they grow roughly, you should be able to work out about how many to seed. Assuming a doubling time of approx 24 hours, you probably would want about 10000 per well to start.