Long-term Live Cell Imaging tips/tricks

[JamesOrth]

I agree in Jennifer, very important to confirm growth with any of these mods. Re the 5% CO2, it should be 5% in the atmosphere of the chamber and local to your cells, not familiar with the chamber but many have a sensor in the chamber with feedback to the CO2 mixer. Are your macrophages proliferating? Playing with density to start with enough but not so much such that your medium is exhausted is a variable that is part of the art of long timelapse…you may have to explore perfusion if possible, adding medium during the run to account for what is lost due to the evaporation issue is not good as the osmolarity of the medium is changed due to evap, now you are adding more medium, but the osmolarity is still off…cells are very unhappy when this happens. Good luck with your experiments!