RE: Freezing Muscle

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From:"Garcia, Vicki {USA~Palo Alto}" <> (by way of histonet)
To:histonet <>
Content-Type:text/plain; charset="us-ascii"

Hi Margaret-
I just started freezing mouse soleus muscle and have tried everything!  The
procedure that works best for me is attaching the muscle to the chuck using
gum tragacanth and then freezing it in isopentane cooled by liquid nitrogen.
I suspend a stainless steel beaker above the container of liquid nitrogen so
that the very bottom of the beaker is submerged in the liquid nitrogen.  It
is critical that your isopentane is cold enough.  The isopentane will begin
to turn white at the bottom of the container and will begin to get viscous.
At this point it is cold enough to freeze muscle.

Vicki Garcia
Roche Bioscience
Tissue Repair
Palo Alto, Ca.
> -----Original Message-----
> From:	Margaret Gondo []
> Sent:	Friday, October 23, 1998 9:39 AM
> To:
> Subject:	Freezing Muscle
> Hi Kids!
> I'm going to be doing some work with muscle(frozen sections)  pretty soon.
> I've always heard horror stories about freeze artifact and things like
> that. I just wanted to know if there is any good reference out there that
> so I can read up on muscle techniques.
> Thanks,
> Margaret

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