I am working on 6 months old mice and would like to find a way to stain all the sensory neurons in DRGs. I have tried the NeuN antibody (MAB377) on DRGs. It works good on 3 months old mice, but not satisfactory on those of 6 months. Maybe the age of the mice affects the staining quality. Is there anybody who has the similar experience or has a better way to stain the somas in DRGs? Any suggestions or ideas would help. Thanks!
If you're looking for a pan-neuronal marker that works very well in the DRG, try Fluorescent Nissl Stain.
I use this in my lab: https://www.thermofisher.com/order/catalog/product/N21483
I was in your position, looking for a good pan-neuronal marker for DRG. NeuN I've found is very weak. Beta-tubulin III is OK, but it seems to prefer larger neurons. Then a friend told me about the fluorescent nissl. You throw it on for 10 mins (1:200 concentration for Far Red) and wash away, just like DAPI. It's really great. It's helped me a lot.
Good luck.
Dear Alex,
thanks so much for your answer! The problem has bothered me so long and your advice is really good. I am going to try it out!
Best
For me the MAP2 worked always the best to stain neurons. However, I have good experience with Tubulin III too. NeuN somehow didn't work for a very long time due to it's unspecificity. I would recommend MAP2. Good luck!
Hi everyone, I have a follow-up question. I prepared dorsal root ganglion culture and cultured for 28 days in the presence of glial inhibitors. I didn't get many big round cells with processes growing out from the soma; instead I got a lot of smaller cells, bipolar with very long processes. My first though was that those were glial cells or Schwann cells but their processes were really long and I included glial inhibitors in the culture medium from the beginning (I did see more fibroblastic or astrocyte-like cells in the beginning but they gradually died out so I believe the glial inhibitors did work). I tried to stain the culture with Tubulin III and s100-beta, hoping to tell which is neuron and which is Schwann cell/astrocyte. And I ended up getting very nice Tubulin III staining the long processes, at the same time co-localized with s100-beta signals (some were stronger and some were weaker). I still couldn't tell if my culture was pure or not. Can anybody give me any suggestions or advice? Thank you!
Hi Katie, if I understand correctly, you are doing the dissociate primary culture, right? Which growth factors are you using? From my experience, if only NGF been given, there will be already fewer glia cells. For glia-sensory co-culture, both NGF & NRG are required. I have cultured the DRG cells for only 1-2 days, the glia cells tend to have a flat-fan-like pattern and sensory neurons have long and thin processes. Maybe you can check your culture in the early culturing days. For the staining, you can also try NeuN to stain neurons; sox10, GFAP and BFAPB for glia cells. Hope it helps you.
Hi,
I am having similar issues with staining DRGs.
I stained my cells with NeuN, as well as beta-tubulin and overplayed - most cells in my culture are stained with NeuN, which makes me think it may be non-specific? After 1 day in vitro I see a lot of small cells with long and thin process, and only a few big ones with lots of processes around them. Would anybody be able to help me with working out what these cells are? When counting the cells after dissection, should I only count the bigger cells?
Thank you very much,
Amelie
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