I'm trying to stain mouse spinal cord sections, (7 um), with Luxol Fast Blue and periodic acid/Schiff and hematoxylin.  In the literature I've been reading, there are some great examples of a nice pink to red stain of the grey matter, and a nice solid blue stain of the white matter.  

Mine look not so good, (purple),  and I've been struggling for a while with it.  I've tried so many adjustments to the protocol that it would be too long to list here.  But I think the heart of my problem is with LFB, differentiating the grey matter completely with out fading the blue-stained white matter.  Somebody please tell me the secret!    

I've experimented with the LFB alone, and believe that differentiating in lithium carbonate, with or with out 70% EtOH, should clear the grey matter of the stain, and leave the white matter blue or turquoise.  But it only clears it somewhat, and any further attempt to differentiate also quickly fades the white matter.  Any help is greatly appreciated. 

Thomas


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