Hello! I am looking for stains that can help me visualize the layers of the cerebellum. My goal is to determine quantity of Purkinje cells, granule cells, and Bergmann glia, and integrity of the folia/layers (for instance, to look for the presence of vacuolization. The cells used will be from frozen and cryoprotected cerebellum slices.
Anyone have any ideas? I am fairly new to all of this, so I appreciate your patience if I've accidentally left out any key info
You can use Nissl stain or H&E stain. You will get a clear differentiation of the white matter, granular layer and molecular layer.
Demos Kynigopoulos Thank you! Do you know what exactly the Nissl and H&E stains stain?
Hailey Rosenfield you can perform immunohistochemistry/IHC using Calbindin for the Purkinje cells marker and GFAP for the astroglia marker, with fluorescence secondary antibody. You can also use conventional IHC, but you need to add a nickel soln. to differentiate the antibody. It will be difficult to observe the dendrite arborization if you use Nissl stain or H&E stain. You can find the protocols in Pubmed or Google Scholar. Good luck.
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