[Histonet] Re: Mouse Lung Fixation

[Baowei Peng]

Dear David,

I\'m wondering if I can use OCT instead of formaloin to inflate mouse lung for frozen sections as decribed in your protocol.
Thanks a lot!


----- Original Message -----
From: "David Weil" <DWeil <@t> ciit.org>
To: <histonet <@t> lists.utsouthwestern.edu>
Cc: 
Subject: [Histonet] RE: Mouse Lung Fixation

 
Dear Andrew

Investigators here have had very good results inflating mouse lungs for
both histology and immunohistochemistry endpoints using this procedure.
It preserves the architecture and causes minimal damage to alveolar
architecture. 

A column of 10% neutral buffered formalin is suspended 20 cm above work
surface using clamp stand. An extension tube is connected from the
formalin column to a blunt 20 gauge needle. The tubing is primed before
inflation so no air passed into the lungs

The mouse is anesthetized with an injectable anesthetic (pentobarbital).
Open the abdominal cavity and then exanguinate by cutting the abdominal
aorta. It is useful reveal the trachea at this time . The thoracic
cavity is cut open carefully to avoid cutting the lungs or any vessels. 

Cut a small incisions in the trachea making sure no to cut it in half
completely. Insert the 20 gauge needle into the trachea and secure it
with a piece of suture. Let the formalin flow into the lungs until fully
inflated. Tie off the lungs with a piece of suture so no formalin
escapes. 

Remove the trachea, heart, thymus and lungs and place in a cup of
formalin for 48 hours. Switch  the lungs to 70% ethanol until ready for
gross trim and processing. 

I do not know how cervical dislocation will affect this procedure, but
we have had very good results with this method and it gives the
investigators reproducible results. If you have any further questions
please contact me. Good luck with the project. 

David S Weil  BS, HTL 
CIIT Centers For Health Research
6 Davis Dr.
RTP, NC 27709-2137
(919) 558-1265
dweil <@t> ciit.org
  
  

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Baowei Peng
1954 Huashan Road
Pharmacy School
Shanghai Jiaotong University
Shanghai, 200030
China
Ph:86-21-62932108
E-mail:pengbw <@t> sjtu.edu.cn