I am a bit new to neuronal tracing and I want to use BDA for some frontal cortex areas of the rat brain. I want to use BDA conjugated with fluorescein, since the microscope I have in the lab is not really efficient with red spectra. I've noticed that most researchers use red staining (Texas-Red if I am not mistaken), but I would like to avoid that. Has anyone done similar experiments with this green BDA? Are there maybe any other commercial BDAs with other colors, that I'm not aware of? (Only blue cannot be suggested since I am using DAPI for nuclei as well).
Yes, fluorescein dextrans are good. Just to explain: The BDA (that you are using) stands for "Biotin-Dextran-Amine", meaning it's a dextran that is not fluorescent but instead conjugated to Biotin. Thus, you can detect it using a molecule that binds Biotin (typically Streptavidin or Extraviding), itself coupled to a fluorophore of your choice (Blue, green, red, infra-red, plenty are available). In other words, by using BDA, your tracing is not endogenously fluorescent, and you will have to "stain" for it and this will set the "color". The advantage is that you can choose the color when doing the staining, depending on other background fluorophores you may have already present in the tissue (GFPs, etc...), providing that you have the variety of Extravidins in your lab (or buy them, I typically use 488 for green, Cy3 for red, and Cy5 for infra-red). This works brilliantly and offers flexible options. Alternatively, if you want an endogenously-fluorescent dextran, there are plenty available from invitrogen (Life Technologies), that are coupled to a fluorophore instead of Biotin. Most commonly used include Alexa 488 or Fluorescein and TetramethylRhodamine (Red). I tend to prefer the red.
https://www.lifetechnologies.com/search/global/searchAction.action?query=dextran+&resultPage=1&resultsPerPage=15&autocomplete=
Best of luck ! ;)
Can you suggest a publication where I can check the data obtained with a 488 Amine on neuronal mrphology?
Cheers and thanks,
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