I need to inject an antibody into the mouse's brain and record synaptic currents from brain slices 6 days after the injection. Is there any innocuous fluorescent molecule I could include in the injection solution that shows me in the slice where the antibody was placed? Thank you!
Hi there,
FM dyes can be used to this end. It has been done in this paper with FM1-43 fx (the fixable version):
https://www.frontiersin.org/articles/10.3389/fneur.2015.00136/full
You can try it, although they waited only 24h for the slicing.
However, check this post
https://www.researchgate.net/post/Long-lasting-non-toxic-dye-injection-in-the-brain
Alternatively, you can refine your coordinates with one or two animals and stain with ink or trypan blue. Afterwards, use the same coordinates for all the animals. If the animals are all of the same age, stereotactic coordinates are quite reproducible.
hope this helps,
Cheers,
J
Hi Daniel,
In the past I used pontamine sky blue (5%) to label recording sites by ionthophoresis. It is very reliable, cheap and easy, works well, and easy to detect with microcopy. For details you can have look at the following publications.
best wishes,
Refik
Article A comparative study of pre-sympathetic and B??tzinger neuron...
Dextran-conjugated fluorescent dyes (e.g. Texas red or fluorescein) are also possible markers. I don’t know that dextrans have no effects on physiology, but doing control injections with only marker and no antibody should be informative, whichever marker you eventually settle on.
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