Maybe this topic was already adressed but I couldn’t find a post about it.
I’m desperately looking for a protocol to efficiently immunostain a small quantity of worms. They are very sensitive so freeze-cracking is not an option, unless you know how to avoid them bursting each time.
I would like to use a finney-ruvkun type of protocol, but on a few (10-50) worms it’s probably useless, I’ll have no worm left at the end…
Freeze-cracking with poly-L-lysine-coated slides is usually fine for a small number of worms. If you have too few of your specimens and are worried about ‘oversquishing’ them, add some animals of a morphological mutant that you can easily distinguish from your worms, like Dpy animals.
Hi, and thanks for your answer.
So you mean, adding a few dumpy for instance will “cushion” the slide and help the coverslip from squishing?
I used superfrost charged slides, worms sticked fine but got squished, so I’ll try your method for sure!
What are you staining for? I assume you are dissecting them to release the gonads, so you are trying to look at the intact worm? I think the previous persons suggestion of using some dpys to help cushion should help you.
I tried to extrude the gonads by dissection, they never come out and/or explode.
Freeze-cracking make them burst, but I might try cushioning with Dumpy worms.
I think doing the finney-ruvkun with an excess of GFP+ worms might help, I’ll try all that thanks!