Re: gluteraldehyde fixation

From:violetta martinez

Hi. I think that you may be missing the exact protocol for this
fixation/preservation of Lac Z enyme . A quick search of the literature turned
up this link which has a probably better/ more complete proocol for the fixation
you require. Apparently this X enyme is the same.
Good luck adn I hope this link helps you.
http://fruitfly4.aecom.yu.edu/labmanual/14b.html

Violetta
Pathology, LHSC

"Gehan, Loralee" wrote:

> Hello,
>
>              I need some help with glut fixation.  My experience with
> gluteraldehyde is that it is a very very slow fix and it needs to be
> refrigerated.  Also I haven't fixed anything in it for more than 24 hours
> and the specimens were very small.  Researchers in my lab are insisting that
> I fix their very large specimens (adult rabbit spine) in .25% glut in pbs
> for about five days.  Then EDTA decal (10%) for two weeks.  I then paraffin
> embed these samples or cut frozen sections.   They insist that this will
> preserve lac Z enzyme.
>               What are your opinions on this?  All of my experience tells me
> that the inner part of the spine will not fix before necrosis sets in.   Has
> anyone every done this?  If you have what was your protocol for fixation?
> Thanks all in advance.
>
> Loralee Gehan
> University of Rochester
>
>




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