I am using HeLa cells for imaging, but after some time I'm working with microscope their morphology change, I do not if they get cold and shrink or what. Because the previous batch of HeLa cells i used to work with them have not had such a problem.I do not know what are the circles around the cells. I appreciate if anyone can help.
Is there a correlation between the cells that are blebbing and those that are expressing your GFP construct. Probably high expression levels of it are making the cells sick. Back off on transfection reagent/DNA concentration.
Christopher B O'Connell . Yes, you're right it is more severe with transfected cells. Thanks for your help. Currently, I'm using GenJet as the transfection reagent, and I follow the manufacturer precisely. Which is 3:1 ratio for transfection reagent:DNA. Do you think I should change the ratio or use less ug of DNA?
Thanks again,
I would try a number of conditions of different concentrations and ratios. From the images, it looks like there may be a misfolding problem where it is all being aggregated. If lower dilutions still give aggregation, the construct may not be able to fold properly and the GFP portion is causing it to mis-fold. You could try putting it on the other terminus.
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