[Histonet] processing mouse lung for cryostat sectioning

[Wadsworth, Marilyn P]

	I am looking for information relating to the preparation of inflated mouse lung to be used for cryostat sectioning. Currently, the protocol being used uses PBS to inflate the lungs which are subsequently tied off and embedded in TBS or OCT and snap frozen in 2-methylbutane cooled in liquid nitrogen. These lung samples need to be unfixed and are being sectioned at 8 microns with disposable knives at a cryostat temperature of -32C.  Any warmer and the TBS or OCT sections, but the lung doesn't. I am suspecting that the PBS inflation may be playing into this - has anyone inflated with agarose, sucrose (or something else) and sectioned successfully? Other ideas? With appreciation...

Marilyn Wadsworth


Marilyn Wadsworth
Microscopy Imaging Center
University of Vermont