Hi all,
I’m trying to pick freshly hatched L1s to count corpses. It’s a challenge to pick sufficient numbers of L1s and to not have them disintegrate before hitting the pads etc. Can someone point me to some hints/tricks to solve this problem. Once the worms are under the scope all is well…it’s just getting them there. I was wondering about washing them off of an egg plate periodically, maybe picking embryos to pads and letting them hatch on there etc. Any help would be greatly appreciated.
Still looking forward to your responses, but I tried a ‘wash’ approach. I put bleached eggs on a plate, waited for a time to get hatchers, washed them off with azide, visualized. Worked great!
I think just taking a gob of them and breaking up the bacteria in the droplet works well enough.
see forums poster Hillel’s protocol: https://www.nature.com/articles/nprot.2007.93
I’m glad you’ve found a method that works for you, and I’m not sure I can explain your initial problem. It should be feasible to pick any number of L1s while harming few or none of them. Your problems may arise from different methods of picking worms?
As Snug suggests, I would recommend having a pick with a flat, smooth shape on the end, flaming it and letting it cool (in the air or by touching it to a bare patch of agar), then getting a glob of bacteria on its lower surface by gently dragging it angled along the surface of the line, much as you might use a putty knife to smooth plaster. This glob can then be gently dragged towards you along the surface of the lawn or the agar gathering up in its path any worm you want to pick up, rather than pressing down on them. Worms can be released from the glob onto an agar pad by gently swirling it into a puddle of your chosen anesthetic (eg azide) without the glob ever directly contacting the surface of the pad; more gentle swirling can disrupt bacterial clumps and distribute the worms on the pad.