Hello all,
I encapsulate human mesenchymal stromal cells within polymeric hydrogels, with degradation enabling encapsulated cell spreading. I have long used Calcein AM to stain and confocal image the encapsulated cells. I now have reason to also stain nuclei, and added 1 ug/mL Hoechst 33342 to the stain solution. Interestingly, I see bright nuclei that are associated with dead/compromised cells, but none associated with live cells. See the attached example image, which is both channels of a confocal z-stack combined. There was no Hoechst signal anywhere in the cell cluster, even at higher gain.
I am thinking that since there is only nucleus signal for dead or damaged cells, there must be some interaction between calcein and Hoechst inside the cell preventing the latter from reaching the nucleus?
I will next try fixing the cells and staining with the same Hoechst alongside rhodamine-maleimide.
Thanks!
- Badges
- Science topic
- Similar topics
- Biological Science
- Histology
- Fixatives