[Histonet] questions re: fixing in general and Histochoice in particular

Geoff McAuliffe mcauliff <@t> umdnj.edu
Wed Mar 25 08:35:32 CDT 2009


Jacqui Detmar wrote:
> Hi all.  Having a bit of an internal debate here, so I would like to get
> the opinions of some of you in Histoland, please.  Here are the
> questions:  
>
>  
>
> 1.        When fixing with 10% NBF, for how long should you fix and what
> volume ratio of fixative:tissue should you use?  
>   
48 hours is a minimum according to R.D. Lillie,  one week is better. 
Really! Thickness of the tissue does not matter, formalin fixes slowly. 
Of course, if you are doing immuno you may need to sacrifice some 
fixation to retain antigenicity. Depends on the antigen.
10X the volume of the tissue.
> 2.       At what temperature should one be fixing tissues?
>   
Room temperature. There is more than ample scientific evidence that 
fixing cold just slows down the already slow fixation process.
>  
>
> Regarding Histochoice:
>   
I never use proprietary fixes, you don't know what is in them and the 
manufacturer can change the formulation any time he/she wants.
>  
>
> 1.        For how long should you fix the tissue?
>
> 2.       What volume ratio of fixative:tissue should you use?
>
> 3.       How long can you store Histochoice-fixed tissues in 70%
> ethanol?
>
>  
>
> I think that's about it.  Thanks in advance,
>
>  
>
> Jacqui
>
>  
>
> Jacqui Detmar, Post-doctoral Fellow
>
> Samuel Lunenfeld Research Institute
>
> Mount Sinai Hospital
>
> 25 Orde Street, room 6-1001AJ
>
> Toronto, ON   M5T 3H7
>
>  
>
> Email:    detmar <@t> lunenfeld.ca
>
> Phone: 416-586-4800 x5607
>
> Fax:        416-586-5993
>
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>
>
>   
Geoff

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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
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voice: (732)-235-4583 
mcauliff <@t> umdnj.edu
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