I was wondering whether there is anyone else have this problem? As shown in the images, almost all the cells are stained with EdU, some are whole nuclei (circle 1), some are puncta inside (circle 2). Is it because all my cells are proliferating? The images are taken with using Nikon confocal machine, Z-stack, 0.25 um per step, 15 steps. Looking forward to your answers, many thanks.
Hi Jun,
EdU staining indicates that your cells are proliferating, since EdU was incorporated in newly synthesized DNA. The % of Edu positive cells depends for how long you have kept the Edu in culture. Usually cells are incubated with EdU for 2-4 hours (https://www.abcam.com/edu-assay--edu-staining-proliferation-kit-ifluor-488-ab219801.html#:~:text=EdU%20Assay%20%2F%20EdU%20Staining%20Proliferation%20Kit%20(iFluor%20488)%20ab219801,flow%20cytometry%20or%20fluorescence%20microscopy). I usually stain my cells (medulloblastoma cells, doubling time 24h) with Edu for 2 hours and obtain around 50% of cells stained.
The fact that in some cells the nuclei is all stained with EdU and in other there are just some dots it depends on the stage of DNA replication (full: early S phase, dots: late S phase)
- Badges
- Science topic
- Similar topics
- Microbiology
- Assays