Re: [Histonet] Mouse Skin Processing

From:Maxim Peshkov



Igor Deyneko wrote:
> I have been havins some problems with processing
>  mouse skin. The blacks are difficult to section,
> then tissue starts flowing away from the
> paraffin and  the sections do not retain
> Hematoxylin stain. That suggest to me that the
> processing and paraffin infiltration were poor.
> Here's the processing protocol I used for pieces
> no larger than 1cm^2.
> 70 % Alcohol - 1.5 hr
> 70 % Alcohol - 1.5 hr
> 70 % Alcohol - 1.0 hr
> 80 % Alcohol - 1.0 hr
> 95 % Alcohol - 1.0 hr
>  95 % Alcohol - 1.0 hr
> 100 % Alcohol - 1.5 hr
>  100 % Alcohol - 1.5 hr( Due to machine only
> having 2 stations for that alcohol)
> Xylene - 1.5 hr
> Xylene - 1.5 hr(Due to machine having only 2
> stations for Xylene)
> Paraffin - 1.0 hr
> Paraffin - 1.0 hr
> Paraffin - 1.0 hr

Igor:
We don't have mouse tissues, but for hard human
tissues (bone, uterus, fibrous samples, skin etc)
have had a similar problem, until start processing
with isopropanol and mineral oil. We doing
processing manually and now forgot any problems
with processing.
All these tissues contain too small volumes of
water and dehydratation for they must be shortened
as mentioned above. Moreover, tissues in
isopropanol will not harden as in ethanol. Mineral
oil is also very  soft clearant than xylene and
others aliphatic hydrocarbons. If you want try
this method, please let me know. I think that it
will be work great for your tissues.
Maxim Peshkov,
Russia,
Taganrog.


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