I'm doing a protocol in which I plate my cells in a 12-well plate with complete medium (i.e. DMEM/F-12 WITH 5% FBS), let them reach ~80% confluency and then I change the complete medium for a starving medium, containing only 0.5% of FBS. Right after I change, some of the cells suffered from alteration in their morphology. It didn't happen in all the wells, but in most of them. I took pictures from moments before and after changing the medium. This cell are EA.hy926, an immortalized endothelial cell.
What can cause this kind of alterations? It seams like the cells are shrinking, although I've never notice anything like that before in other protocols and other plates. Could it be incompatibility with cell plate i'm using (Corning).
Thank you!
Iy looks more like an optical effect. The top image is Brightfield and not quite in focus, the bottom is almost in Phase. there may be a meniscus effect as well as a change in medium refractive index. Did you readjust the condenser and filters?
IF the cells really are shrinking, they are probably sick and dying, and the starvation medium is insufficiently nutritious (containing only 0.5% of FBS-does this mean that the FBS is the only additive in this solution, or are there any other constituents?)
Ellen A G Chernoff Thanks so much for your answer! I actually didn't change anything in the microscope from one image to another. I used my cell to take these pictures, so I think that can be the cause. When I looked in the microscope directly, this kind of optical effect do not appear.
Abhishek Balmik
I checked again 24h later, and these characteristics persisted, including a great amount of the cells have died.Roger Bayston I also include pen/strep in the medium.
I performed several tests, and I came to the conclusion that it was some kind of hypersensitivity to the medium pH. When I left the medium ~10min inside the CO2 incubator, the problem disappeared.
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