RE: pollen

From: Mellard, David (dam7@cdc.gov)
Date: Mon Nov 16 1998 - 04:33:51 PST


Date: Mon, 16 Nov 1998 07:33:51 -0500
From: "Mellard, David" <dam7@cdc.gov>
To: cp@opus.hpl.hp.com
Message-Id: <aabcdefg3600$foo@default>
Subject: RE: pollen

Back in another life when I did tissue culture for a living, I regularly
froze cells in (if I remember the % correctly) 10% DMSO (dimethylsulfoxide).
This certainly should work for pollen and allow you to store it for years.
The procedure involves sealing the cells in glass ampoules in 10% DMSO and
then slowly lowering the temperature to below freezing. There are expensive
instruments to do this but we did it simply by putting the vials first in
the refrigerator, then in the freezer.

The catch is that after going in the freezer for a few hours, the vials have
to be slowly lowered into a metal container and immersed in liquid nitrogen.
I forget the temperature but it pretty much stops all biological processes,
which is what allows the cells to remain alive. When retrieving the cells,
the vials are thawed quickly in a warm water bath.

The above process should work for pollen. One will probably need to dilute
the thawed pollen with water followed by gently centrifuging and then
decanting the liquid to get back to a solution free pollen.

You just might get by with freezing the pollen in %10 DMSO and keeping it in
the freezer if the liquid nitrogen seems too much. I few trials should tell
you whether or not the pollen remains intact and active.

David
Atlanta



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