Patching Platynereis Dumerilii

[Steffen]

Hi there,
I could need some help with my current project. I'm trying to patch clamp some cells of the hardly 150 micrometer sized larva of Platynereis dumerilii. Unfortunately, the larvae have a very tough collagen cuticle, which is really hard to penetrate. Just poking a hole in it squeezes the animal so much, that cells on the inside get destroyed. Dissolving the cuticle by enzymes didn't work as expected and just killed the animals.
So now I'm in desparate need of a good and quick way to find a way through the cuticle and patch some cells inside. Optimally, it should be possible to use only one electrode. One manipulator is occupied with the holding pipette, with which I fixate the otherwise free swimming larvae. The other one carries the measuring electrode, which btw. gets dirty when poking a hole through the cuticle.
I already tried to stab a hole, remove the electrode, and come back with a clean one. Of course the animals leaks out, when I pull out "the plug" and surely I don't want that.
Does someone have a nice idea, which does not involve laser surgery and nano-bots?

[Steffen]

If there is someone, who is interested:
Up to now it seems like the best way actually is laser surgery. To shoot a hole into the cuticle is easy. Afterwards finding it is not so easy, but there are some ways to get hints. For example, there are one or more bubbles of vaporized or burned tissue, which indicate where appr. the hole might be. With enough positive pressure, one can search along the surface of the larva without getting dirt onto the electrode...