XENOPUS LATERAL LINE VISUALIZATION

Procedure

To see the lateral line in Xenopus:

1. Use older embryos, like st. 45-48
2. Do not use gent! Gentamycin and neomycin kill the neuron cells.
3. If seeing live, do this:
   1. Stain w/ FM1-43 dye for 7 minutes (not longer)
   2. Wash in plain water (not MMR)
   3. View on fluorescent scope.
   4. If using the fixable form of the dye, fix it, but in my experience, it doesn’t actually last past immunohistochemistry…
4. If doing immunohistochemistry with a marker (like the NompC channel antibody from Stefan Gruender):
   1. Make sure to bleach with peroxide/MeOH
   2. Doing Alk-Phos/fluorescence doesn’t work well if you add primaries and secondaries together. Do the AP detection, post-fix, and then do immuno with fluorescence. Or, just do it with 2 different fluorescent colors.

This protocol is from Levin laboratory under Dr. Michael Levin at Tufts University.

PROTOCOL DEX