Hello Ameena,

what is the aim of the DAPI staining? and the type of your cells?

If you need only to mark the nuclei and take photos, I advise you to use sterilized square coverslip. You can put them inside of your 6 well plate and when the staining is done, you can flip and mount the coversplip on the slide. I don't know which type of cells are you working on, but, in case, you can cover the coverplip with poly-d-lysine before seeding the cells. Another possible way is to use ibidi slides. They have a special insert (like 8 mini wells), so you can seed less cells and treat them, at the same time, with different drugs or eventually different antibodies.

The staining with the Dapi is quite simple. I usually use Dapi at the end of my immunofluorescence experiment to mark the nuclei. I dilute my dapi stock in PBS and treat my samples for 5' in the dark. Then I wash with PBS 1X 2-3 times to remove the excess and finally, I close the slide with the mounting media.

I hope that my answer can help you!

@Martina Vincenzi Thank you mam. The cell line to be used is esophageal adenocarcinoma, basically an adherent cell line. The objective is to assess how the drugs (at IC50) affecting the nucleus of these cells. If possible, can you recommend any articles based on the assay?

DAPI is best if the cells are fixed and permeabilized. Label with a very small concentration, just 0.2 ug/mL for 5-10 minutes in PBS or other physiological buffer at RT.

If you wish to do live cell labeling, then I recommend instead using Hoechst 33342, which is more cell permeant, labeled at 0.4 ug/mL for 5 minutes in media or suitable live cell buffer. Be aware that nucleic acids like Hoechst will affect DNA function, such as proliferation, so isn't recommended for culturing or long term imaging after label; only for end point assays.