[Zbrafish] Lipophilic tracers and MeOH preserved embryos

Chi-Bin Chien via zbrafish%40net.bio.net (by chi-bin.chien from neuro.utah.edu)
Mon Jun 22 03:33:13 EST 2009


Rehydration will not help--the problem is that lipophilic dye tracing  
in fixed embryos depends on the fact that aldehyde fixation (like PFA)  
crosslinks proteins but leaves plasma membranes (like axonal  
membranes) intact. If you extract lipids, they are gone and not  
retrievable.

Chi-Bin

On Jun 19, 2009, at 11:17 AM, Cameron Wyatt wrote:

> I have recently received a batch of 3 days post fertilisation
> zebrafish embryos in which I had intended to trace the optic
> projection with DiI or DiO. Unfortunately the embryos were preserved
> in MeOH.
> Can anyone direct me to a protocol for rehydration which would make
> the embryos suitable for tracing or has the MeOH rendered them
> unusable?
> Any suggestions will be greatly appreciated.
> ~ Cameron
>
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