[Histonet] Golgi-Cox & aqueous mounting media

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I am trying to get the golgi-cox stain working in rat neonatal brain tissue.  I am using perfusion fixed tissue (10% formalin) and then immersing in impregnation fluid.  I am using mixture taken from Ramon-Moliner's 1970 paper in contemporary methods in neuroanatomy.  I do not have access to the equipment used for plastic or celloidin embedding and thus, am sectioning the tissue on a freezing microtome after washing and cryoprotecting in 30%sucrose.  I am finding that it is necessary to cut sections at least 180um thick to get decent sections.  Otherwise, the sections fold upon themselves and are not unfolding.  

So, is this what most people have found trying to cut frozen sections that were treated with the Golgi-Cox impregnation fluid?  And what do I do to coverslip these thick sections?  What is Canadian Balsam?



Also, I have been using hardset from vector to coverslip slides from a seperate experiment that were stained with fluorojade.  So far I don't like this aqueous mounting media.  It seems to thick and bubbles form far to easily.  Can I dilute it with PBS?  I used to use Gel-mount which I liked better.





Brad 

Drexel U of Med School

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