Proper cell seeding technique is a critical part of setting up your experiment and helps ensure that your Agilent Seahorse XF data is reliable, reproducible, and accurate.
This video reveals several best practice techniques for cell seeding.
These include leaving the cell plate in the culture hood for one hour after the seeding is complete to help avoid edge effects due to thermal gradients,
and discussion of cell seeding density (cell per well).
Pre-incubation of the cell culture microplates with the newly seeded cells at room temperature prior to moving to the incubator is an effective technique to control "edge effect" and to ensure even distribution of the cells in each well. Whether you are using adherent or suspension cells, primary cells or cell lines, your Seahorse XF data will benefit from following these best practices.