We stained the mice brain (not fix with formaldehyde) with golgi-cox solution for 2 weeks. However, upon sectioning with vibratome (100 - 200 uM thickness, embedded in 4% agarose gel), the tissue looks brittle and torn during sectioning. We hardly get an intact slice. We did not fix with formaldehyde since some articles did not recommend such step for neuronal staining. Kindly advise.
Hi!
I did Golgi-Cox staining of mouse brains during my master thesis and had similar problems. I used a staining based on this protocol: Article A method for vibratome sectioning of Golgi-Cox stained whole rat brain
I remember that cutting the tissue was difficult and I had to lower the speed and the amplitude of the vibratome. I don't remember the exact settings, but could check in my old lab journal, if I still find that. I also did not fix samples in PFA or formalin, but immersed directly in Golgi-Cox solution. What I remember, is that immersion in 30% sucrose helped with the tissue consistency, although it was still difficult to section.
Hope this helps!
Cheers
Sven
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