[Histonet] Re: Mouse lung fixation

From:Sven van Eijl

At 07:35  10-01-05, you wrote:
>In a message dated 1/9/2005 5:44:15 AM US Mountain Standard Time,
>Andrew.MacDuff@ed.ac.uk writes:
>am just starting a research project and need to fix mouse lungs for embedding
>in parafin so that I can use them for histology and immunohistochemistry.
>Could you please tell me the best way to fix the lungs? So far I have tried
>removing the lungs and then instilling 10% neutral buffered formalin into the
>trachea with a syringe. This seems to work but I'm not sure how 
>reproducible the
>alveolar architecture will be. Also do people exsanguinate the mouse before
>fixing the tissues? (we've been using cervical dislocation and then just 
>the lungs)

Hi Andrew,

Are you going to do any morphometry on the alveoli ? in that case it is 
important that the lungs are always fixed at a set pressure. We take the 
lungs out of the body and attach them by the trachea to a set-up in which 
we keep the column of fixative which is pressurizing the lungs at 25 cm 
(through the communicating vessels principle). This way you can ensure that 
each lung is inflated to the same extent. We use Carnoy's, which acts very 
rapidly, but also seems to leak through the tissue quite a bit once it is 
fixed, probably because it is chloroform-based. Personally, I rather not 
use cervical dislocation, afraid it might damage lung structures, but I'm 
not sure that's really an issue.

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