Hi, I'm currently trying to perform an in vitro Live-Dead assay using HUVECs. Unfortunately I'm encountering trouble visualizing the stained adherent cells with the fluorescent microscope, most likely due to the hydrogel coating the bottom. I usually coat the wells first before adding the cell suspension which I assume diffuses through the gel to the plate surface to adhere. Does anyone have any insight on how I might test the effects of the hydrogel using this method?
Also Alamar blue seems to not work as none of my hydrogel plated samples were reduced probably due to inability for the dye to be taken up due to the hydrogel obstruction.
Thanks for your time
Dear Brandon Applewhite,
You can use cover slip based method by coating with the hydrogel and seed the cells. Then, you can proceed for the staining prior to observation under fluorescent by inverted the cover slip. Better results would be obtained later on. Wish you best of luck!
Fauzi Mb Thanks for your response. Do you mean to place the cover slip within the well?
Brandon Applewhite yes, you have to keep the cover slip in the well. Once cells are attached to the coverslip (coated) you can do staining and when you want to see under the microscope invert the coverslip.
- Badges
- Science topic