We attempted a series of colony forming assays of cancer cells for irradiation. However, due to some vibration issues from our incubator, the cells settled towards the center of the plate following our plating and prior to irradiation. The cells were plucked from a cell culture at different evolutionary stages so we don't have a way to go back and do it again.
Is there any way to salvage data from this in a scientifically acceptable/rigorous way? Is comparing colony area to 0 Gy samples a valid method? Ideally we'd like to find cell survival fraction, but any other useful data we could get from this would be awesome. They've already been fixed and stained.
Hi John all I can think of is that you abandon the "clonal" bit of the colony forming assay and just go for the population doubling (PD) between each condition.
If you know the number of cells seeded at the start of the experiment (plating) and the end (fixed and stained) then the PD = (log (harvest no/seeded no))/log2
You can get the total cell number at the end by using a nuclear stain on the cells (haematoxylin of Dapi etc) and either manually counting or automatically using i.e. an ImageJ batch command.
Obviously any measurement (such as total area) is also valid if enough replicates of each condition has been performed and shows consistency.
Sorry I can't be more help!
Gary
- Badges
- Science topic
- Similar topics
- Physical Sciences
- Materials Science
- Plating