centrifuge of spores

[Luigi Meneghelli <meneghel@AVA.BCC.ORST.EDU>]

Wim Vanhauwaert
    Sorry about the mis-understanding, I was trying to be brief in my description.
I don't know if you can use this method to separate different size particles, unless
you fine tune the amount of centrifugation or solution (like a density gradient).
However it is effective in separating heavier spores from leaf parts which generally
float.
    Start out by putting your spores into a centrifuge tube and mix with your bleach
(sterilizing) solution and vortex, shake... then use the centrifuge to get them out
of solution, by pelleting them at the bottom of the tube, pour off (or remove with a
pipette) the sterilizing solution, and pour sterile water into the tube.  Re-suspend
the spores into the sterile water and rinse bleach off spores by shaking or
vortexing.  Wash the spores with sterile water a few more times (spin down, remove
water and then resuspend).  Finally after three rinses resuspend spores in a small
volume of water and while in suspension drop the water and spores out onto a
germination plate.  This last step allows you to spread the spores out evenly on a
petri plate with germination media (or just filter paper).  The drop of water will
be absorbed by the media or evaporate leaving the spores evenly spaced.
    My other suggestion was for sterilizing spores if you don't have access to a
centrifuge.  Because of their small size spores may not stay in cheese cloth, which
is commonly used.  Camera lens cleaning paper is a really tight weaved polyester (my
guess) type material that will contain the spores while letting the sterilizing
solution clean the spores.  Fold the spores up in the lens cleaning paper close
witha paper clip and place in bleach, wash then transfer to sterile water.  After
sterilizing smear the spores out on your germination plates.
I hope that clarifies things for you.

Luigi Meneghelli
Oregon State University
Dept of Horticuluture.

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