I have been trying to measure lung epithelial proliferation in vivo during/after an influenza infection and i have been having a trouble picking up any staining at all.
We have been using the EdU click-it kit for flow cytometry (APC staining) and I have been able to pick up staining in vitro, however I have not been able to get it to work in vivo and have not actually been able to pick up any staining at all for the EdU kit.
We normally inject the EdU i.p. in our mice at 25 mg/kg. At first I thought perhaps there just wasn't very much epithelial proliferation at the time we were looking at so I have tried a pilot looking in the thymus and staining for CD4 and CD8 T cells as I would expect to see a great deal of proliferation there anyways. I injected the mice and took the thymus 24hours after EdU injection and was still not able to pick up any staining at all.
In the literature there are a few groups that have done EdU staining in vivo before and I'm at a bit of a loss as to why I am unable to get it to work. If anyone has any suggestions, advice or ideas that I could try they would be much appreciated.
Hi Kristy, the small intestine will for sure have picked up plenty of EdU with your dose/time point so it would make a good positive control for the in vivo EdU incorportation. Also, is the scope for in vitro the same one you are looking at the in vivo tissue section slides on so you know it isn't a scope/excitation issue with the far red fluor?
Follow This Link please:
https://webpage.pace.edu/ms16182p/troubleshooting/mice.html
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