Cleaning worms..

David H. A. Fitch david.fitch at nyu.edu
Thu Jul 11 14:03:37 EST 2002


Hi Paul,

sometimes the simplest methods are the best.  This one is from field 
nematologists and is called the "Baerman funnel".
Attach a rubber tube to the end of a funnel and clamp it closed with 
a pinch-clamp or screw-down type "C" clamp.  in the mouth of the 
funnel, put a small screen, which acts as a support for a filter. But 
instead of filter paper, use two KimWipes.  Wet the KimWipes by 
squirting water into the funnel, and remove the air bubbles that 
invariably get caught in the tubing (e.g., by squeezing the tubing 
several times or opening the clamp to let some water down into the 
tube).
Put your worms + agar (or worms that have burrowed into agar, or 
worms in dirt, or live worms in any matrix) onto the KimWipes.  Add 
water to just cover them.  Over a period of a couple hours, the worms 
will crawl out of the agar pieces and through the mesh of the 
KimWipes.  They will then sink to the bottom of the clamped tubing. 
Collect the worms by opening the tubing slightly and letting a few 
drops of water drip into a test tube.  Your worms will have no agar 
or dirt.

Another method is to float the worms on 30% sucrose and centrifuge. 
Agar will generally pellet faster than the worms.  But the worms are 
all shriveled up.

Dave

>I am a researcher currently attempting to perform in situ
>hybridisation experiments on adult C. elegans.  Unfortunately, the
>experiments are being hampered somewhat by the presence of agar (from
>the growth plates) in the reaction mixture.
>
>Can anyone give me a RELIABLE protocol for removing agar fragments
>from the worms?
>
>Thanks a lot,
>                     Paul McVeigh
>                     Parasitology Research Group
>                     Queens University Belfast

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