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silver staining (dendritic spine visualization) protocol?


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#1 leevonk

leevonk

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Posted 04 July 2009 - 08:06 PM

Does anyone have a silver staining protocol for visualizing dendritic spines with light microscopy? This is one I found at (http://synapses.clm....e/visualize.stm), any alterations or important notes anyone could suggest? I mainly do extracellular in vivo cortical recording and stimulating, not much experience with staining.

The classical Golgi impregnation method is very elegant in its simplicity:
1. Immerse a block (approx. 10x5 mm) of formol-fixed (or paraformaldehyde- glutaraldehyde-pefused) brain tissue into a 2% aqueous solution of potassium dichromate for 2 days
2. Dry the block shortly with filter paper.
3. Immerse the block into a 2% aqueous solution of silver nitrate for another 2 days.
4. Cut sections approx. 20-100 Ám thick.
5. Dehydrate quickly in ethanol, clear and mount (e.g., into Depex).

thanks!




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