[Histonet] Nuclear DAPI staining in frozen sections of bones

[Park, Shin-Young]

I am doing nuclear DAPI staining in frozen sections of fresh, non-decalcified mouse femurs.
The fresh femurs were snap frozen in OCT block by 2-methylbutane (isopentane)/dry ice.
Cryosection of the bones were done by using the Instrumedics CryoJane tape transfer system.
After transferring sections to slides, the slides were kept in -80oC until staining.

But I have problem with the quality of nuclear DAPI staining, although H&E staining is pretty good.
The nuclear DAPI staining (Vectorshield with DAPI or 1 uM DAPI, 10 min in PBS) is fussy and unclear.
It is hard to identify each individual cell by nuclear staining.

If anyone else has experience on DAPI staining in frozen bone sections, could you share any suggestion to improve the DAPI staining?

Any help would be greatly appreciated.

Shin-Young Park