Hi all,
I'm currently imaging 150um-thick sections of a rat-hippocampus attempting to analyze the number of synapses along basal dendritic branches in CA1 Pyramidal neurons. During my imaging, I'm noticing that there are stained fibrillary material near the synapses. Does anyone have any idea what these might be/how to correct for them in the future?
Here is a quick overview of my staining procedure:
-Using FDneurotechnologies Golgi Staining Kit
1. Animals were anesthesized, rapidly decapped, and brain removed
2. Brains were placed in solution A+B for 2 weeks
3. Brains were then switched to Solution C for 3-7days
4. Tissue was rapidly frozen using isopentane, kept on dry ice for 10min and sliced on a cryostat kept at -24 degree Celsius
5. Tissue was stained 1-2 days following slicing
Additional Notes:
-All tissue was fresh and frozen only before sectioning.
-This isn't occurring at every dendrite but I have noticed it on some of the dendrites of the animals I have looked at.
Thank you all for your time!
Nicholas hello,
For more clarity and optimal results, please, consult this protocol:
www.frontiersin.org/articles/10.3389/fnana.2016.00038/full
I hope this helps :)
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