Hello colleagues, I’m currently working on a traumatic brain injury (TBI) model in rats and focusing on quantifying alive (viable) neurons in the cortical region adjacent to the incision or impact site. However, this area contains a heterogeneous population of cells, including glial cells and possibly degenerating neurons, making it difficult to clearly distinguish and count only the viable neurons.
I’m using Nissl staining to visualize the neurons, but I’m not fully confident that I am accurately identifying and counting only the healthy, living neurons.
My question is: What is the best approach to confidently and reliably identify and count viable neurons in this context? Are there additional staining techniques, markers, or morphological criteria you recommend to improve the accuracy of cell identification?
Thank you in advance for your guidance and suggestions.
I’ve attached images from my sections. I would appreciate any advice on:
Whether these sections looks suitable for neuron counting?
which of these cells are viable neurons?
Hello, I'm not sure how reliably you can detect small neurons with Nissel staining. It would probably be best to do a double staining: 1) valid neuronal marker (e.g., HuC/D), 2) activated caspase 3. HuC/D-positive but activated caspase 3-negative cells should then be viable (or at least still viable) neurons.
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