[Histonet] Dil labeling

[Narjes]

Hello all,
Please I need some help in the lipophilic dil labeling of free floating brain sections. The mice brain were fixed in 1.5% PFA by cardia perfusion, then left one hour in the same fixative. I used the Dil labeling solution for a 200 um thick sections, the dilution was 1:150 in PBS. The sections were incubated for 48h in the Dil labeling solution then 48 in PBS in the cold room in the shaker. The problem that I had a very high background and I could not see even one spine. I need to stain only a few neurons so I can do a reasonable quantification of the size and number of the spines. Does anybody has an idea if in this case the solution or the crystal Dil is better and can I get an optimized protocol for this kind of staining.
Thank you so much
All the best
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