Hi everybody,
I need some advise about a strange pattern I obtained on yeast cells with a DAPI staining
The protocol was the following
On L-polylysine coated wells, the cells were air dried, fixated 30 minutes in 3,7 % formaldehyde, washed three time with PBS, fixated 5 min with methanol-ice cold and 30 sec with acetone ice-cold (until dried)
Then came the staining with my antibodies and final staining with DAPI
One of my slide then showed a diffusing DAPI pattern (whereas all control slides showed a typical nuclei restricted pattern)
given the pattern either it is a diffusion of nucleic acid within the whole cytoplasma of my cells (however a retrocontrol culture on the same showed that they are fully alive) or a staining of something else (either a cytoplamsic or a fungal cell component)
My question is then :
does the DAPI is known to stain other components (sugar links for example if we assume that my labelling concerns the fungal cell) that nucleic acids molecules ?here is a picture of the "problem"
