[Histonet] Fwd: Help in Dil labeling of brain sections

[Linda Margraf]

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Begin forwarded message:

> From: narjes baazaoui <baazaouinarjes <@t> gmail.com>
> Date: May 14, 2014 at 1:35:04 PM CDT
> To: histonet-owner <@t> lists.utsouthwestern.edu
> Subject: Help in Dil labeling of brain sections
> 
> 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
> Narjes
> 
> -- 
> Narjes Baazaoui, PhD student
> Institute of Basic research (IBR),
> Laboratory of chemical neuropathology
> 3476715743