RE: Mouse lung processing

From:Susan Bell


Here's a good way for when you're "learning" that I find easier that 
how I used to perfuse lungs.

Use a 3 mL syrings with a 22X1 with 1-1/4" mm ball animal feeding 
needle (Popper & Sons Inc.  New Hyde Park NY 11040).  Cut the trachea 
as far up the neck as possible and hold on to it.  Gently slip the 
feeding needle into the trachea and hold the end of the trachea with 
forceps while gently injecting 10% neutral buffered formalin into the 
lungs.  You will be able to see the lungs inflate.  Still holding the 
end of the trachea gently remove the lungs from the animal and put in 
jar of formalin.  It's very important not to touch the lungs.  It is 
possible to do this entire procedure without handling them at all.  I 
like the feeding tube better than a regular syringe needle because in 
reduces the risk of puncturing through.

I'm currently using GFP (Green Fluorescent Protein) as a marker in a 
study and don't let the lungs sit in the formalin for more than about 
24 hours before having them processed.  According to different 
studies, GFP doesn't hold up well in formalin for longer amounts of 
time.  I haven't done any studies on frozen section, so I can't give 
you a comparison.

I hope this helps.

Susan Bell

At 11:56 AM -0400 9/3/02, Prodhan, Parthak,M.D. wrote:
>Hi Histonetters,
>I am new to this field of histology. I was hoping to get some help from the
>group members for my queries.
>1. How does one fix and embedd postnatal mouse lung tissue? Stress on
>maintaining architecture of the alveoli. I have had problems in maintaing
>architecture with frozen sections. Any suggestions are most welcome.
>2. For immunoflorescence on mouse lung sections using paraffin 
>sections. Does it
>give better results as compared to frozen sections.
>Thank you in advance
>Parthak Prodhan
>Pediatric Pulmonary Lab
>MassGeneral Hospital for Children
>Boston, MA


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