[Histonet] Help with histo on fish eggs

[nizar limaiem]

Dear doctor

I would thanks you to your rapid replay.

I have seen an article in journal of developpement 2001 about the
histological study of the

developpement of oreochromis niloticus , i am very interesting, so i hope te
done a similar work aplied to a fish species dicentrarchus labrax L. in this
artical there are no protocol indication.

I have applied the protocol witch given me, i rplaced the xylene by toluene
because we haven't en our lab. I don't anderstend if the 30 min periode is
the time all deshydratation  steps (70°,95 and 3x100%) or for each
concentration. in my protocol i used 30 min for each concentration. when i
use micotome at 5 microns the eggs dont stay in the block as if they are not
prfusd by paraffine. the few section abtened colored with H/E showed
disordered structure thtcould not be interpreted. so I remplaced the toluene
by butanol  and 1:1 xylene parafine by ottix parafine (butanol is not
missible with paraffine) the section obtened showed clear tissu but not
sufficient to be interpeted.

thanks in advence

 I'm afraid may way of expression in nglish are not very good.

Sincerely

limaiem nizar.

Student in marin biotechnology

University of  biotechnology, Monastir, Tunisia