I am performing for the first time a colony formation assay in soft agar with a tumoral cell line and now that my experiment has reached 3 weeks of incubation, I expected to have quite large colonies, however, my colonies are too small to be detected in a gel documentation system. I can see the colonies at 10x even at 4x, and I have plenty of them, so it should be quite difficult to count them at microscope without human error.
Moreover, I stained cells with crystal violet 0.005% and I took a picture exposing with white light and nothing seems to appear.
Has anyone had the same problem? How do you solve it?
Hello Raj, what do you mean by a "good camera"? A camera attached to a microscope or any powerful camera? How do you do?
There is an automated colony counter, the GelCount, which can count colonies from the 25um-30um range and larger. Not sure if that will help you but it is another option. Id say the images on that system in particular are better than most cameras but that is just my opinion. https://www.oxford-optronix.com/en/GelCount
Also, on that page is a publication list at least a few hundred articles deep counting both large and small colonies.
Cheers,
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