[Histonet] Sorry to disagree! Re:Mouse Lung inflation-OCT

[Gayle Callis]

Ann and others,

Sorry to disagree, we do this routinely and never dilute the 
OCT.  Introduction of OCT into lungs is via a v shaped cut on TOP of 
trachea.  The trachea is never severed completely across the lumen or the 
trachea retracts into lung cavity. We use 2 mls OCT with a dulled 18 guage 
needle (we dull sharp point and edges with a fingernail file containing 
various grits sand paper!) inserted into the trachea gently towards the 
lung.  After lung is OCT filled,  gently remove lung from mouse, dissect 
heart away, and snap freeze.  We experience no difficulty nor damage to 
lung morphology.

We used to dilute the OCT with PBS 1:1, but found it was not necessary.

   At 07:29 AM 1/12/2005, you wrote:




>OCT is almost impossible to use to inflate the lungs, it is very viscous,
>but I have used a 1:1 OCT and 5%sucrose solution to inflate adult mouse
>lungs and it adds tremendously to the morphology of the frozen lung tissue
>and makes it easier to section also. Immunohistochemistry and in situ
>procedures have been performed on these samples with no adverse effects.
>
>
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Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)