Re: Paraform/fluoro
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From: | "P. Emry" <emry@u.washington.edu> |
To: | Karen Larison <larisonk@uoneuro.uoregon.edu> |
Reply-To: | |
Content-Type: | TEXT/PLAIN; charset=US-ASCII |
Thanks, really. I told him I reluctantly was going to show this to him.
It is good to know why.
Trisha
On Fri, 3 Mar 2000, Karen Larison wrote:
> Trisha,
>
> What fluorescent retrograde tracers is he using? If he's using DiI, I would
> absolutely recommend sticking with aqueous-based fixatives, preferably
> paraformaldehyde. Don't use Prefer if it contains alcohols or organic solvents!
> Other fluorescent tracers may be less picky, but I would still advise caution,
> because most neurotracers work because they can't breach the membrane once they are
> in the axon, so you don't want to use anything that can possibly perturb the
> membrane. That's why people use paraformaldehyde rather than formaldehyde in these
> applications. The methanol in formalin has the potential to perturb the membrane and
> you can get cross-talk between in axons in the nerve. In otherwords, the dye crosses
> too easily from axon to axon instead of staying within the axon of interest.
>
> Paraformaldehyde is no different than formaldehyde once it's in solution. However,
> it will tend to polymerize over time, and precipitate. Thus, you have to make it
> fresh. The purpose of the methanol in formalin is to keep the formaldehyde in
> solution. But it can and does mess up cell membranes.
>
> Personally, I wouldn't ask him to venture into places unknown with unknown additives
> in the commercial fix. It's just too likely to mess up his experiments. And some of
> these tracer experiments are quite long and involved. He should stick with the
> paraformaldehyde.
>
> Karen Larison
> Institute of Neuroscience
> University of Oregon
>
>
>
>
> Date: Thu, 02 Mar 2000 18:46:52 -0800 (PST)
> From: "P. Emry" <emry@u.washington.edu>
> Subject: Paraform/fluoro
> To: HistoNet@pathology.swmed.edu
>
> Hi all,
>
> I am working with a post doc who wants to inject some "fluorescent
> retrograde neurotracers" in to pigs and use paraformaldehyde to fix the
> specimens. (Forgive me if I have muddled that up.)
>
> He is being very kind in considering my formaldehyde aversions and
> said he would work after I go home. He is reluctant to use my
> substitute Prefer thinking it may effect the fluorescent signal.
>
> Have any of you worked with Prefer or any other
> formaldehyde-paraformaldehyde substitute for that use? (I have never used
> paraform. and have no idea how it differs from formaldehyde.)
>
> I am sure he will want some references if possible.
>
> Thanks,
>
> Trisha
>
>
>
>
>
Trisha
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