|From:||Roger Moretz <email@example.com>|
Since I am officially "on vacation" this week, I am
rather late about getting to reading any email. So,
even tho' I am nowhere near that dear vibratome, a
couple of additional comments. Granted that 10 to 20
microns is very difficult. 40 microns is easy with
fixed tissue (such as the perfused brain, or just NBF
fixed liver--about 2mm thick sections collected at
necropsy). I don't do this a lot, so I cannot
remember knife angle or speed or amplitude. The
critical parameters for me have always been to ensure
that the bath remains chilled (mine has a Peltier
cooler that requires a high capacity chiller for a
cold water source, and even then I routinely add ice
to the bath. For brain, liver and kidney I prefer a
#22 scalpel blade. The blade can be mounted to use
the straightest portion of the cutting edge. I find
these to be far superior to single or double edge
blades. Haven't tried sapphires--too expensive for
the limited use we have. Finally, the practice is so
important. Due to the long times between use, I find
that I spend 45min to an hour just re-familiarizing
myself, and then getting into a consistent rhythm. I
have yet to get decent sections of skin off the bloody
thing. Tried for nearly two months. Not even
reasonable results at anything less than 75 microns.
Just about everything else can be done. Best of luck!
Dept of Toxicology
Boehringer Ingelheim Pharmaceuticals
--- Valleygal@aol.com wrote:
> I have recently -- very recently -- learned about
> cutting vibratome sections
> and I totally agree that 10-20 micron sections would
> be tough. I did 40
> microns with great success on perfused brain,
> Before I started I called Ted Pella and was advised
> to practice on
> hard-boiled egg white. So off to the cafeteria I
> went to buy an egg and amid
> lots of cracks about egg salad and the like I spent
> an afternoon practicing
> and the next day I cut that brain like a pro! The
> consistency of hard-boiled
> egg white was very similar to the perfused brain.
> Practice. Yes, do practice.
> Andi Grantham
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