[Histonet] RE: mouse testis in Bouins

[Frances Elizabeth Barron]

Hi Margaret,

Our protocol for whole mouse embryos E14.5-E18.5 was to fix in Bouin's for 5-7 days at room temp (I have gone longer, but it isn't exactly recommended). Most of the length of time, however, was to compensate for the large tissue size and need for good penetration. I'm not sure how that converts to your particular tissue of interest.

For long term storage, John Shelton at UT Southwestern (who did our vacuum processing for large embryos) told me that it was preferred to put them in 1% neutral buffered formalin and store them at room temp. We had previously been storing them in 70% EtOH, but John said that the long exposure to EtOH leads to excessive drying of the tissue and ultimately brittleness if used later. I'm assuming this thought could be applied to any tissue piece, but I don't have enough experience to really know. We have successfully gotten beautiful paraffin sections from 3mo-1year samples that have been stored this way. 

I'm hoping this will be of some help to you, and perhaps others in the list can comment.

Best of luck,
~Francie

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Francie Barron, Ph.D.
Postdoctoral Fellow, Joseph Wu Lab

Stanford University School of Medicine
Lorry I. Lokey Stem Cell Research Building
265 Campus Drive, Room G1105
Stanford, CA 94305-5454

Phone: (650) 724-5564 or (650) 724-9240
Fax: (650) 736-0234

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Message: 7
Date: Fri, 14 Sep 2012 10:06:33 -0300
From: "Margaret Horne" <Mhorne <@t> upei.ca>
Subject: [Histonet] mouse testis in Bouins
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <505301A9020000D10001841D <@t> oes-grpwise.novell.upei.ca>
Content-Type: text/plain; charset="us-ascii"

 Hello Everyone, I am asking this for a friend.
 
How long can mouse testis be kept in Bouins without distortion of cell
morphology? Days? weeks? months? years?
 
I noticed in the Archives that many people fix in Bouins , rinse, then
store in 70% EtOH. This is preferable I assume. Again, how long is ok?
 
                                                        
   Thanks in advance for the sharing of your accumulated wisdom, 
                          Margaret