96-well liquid culture lifespan

Model Organism: Worms Methods & Reagents
1

Hi, is it a good idea to shake worms at 900 rpm while doing lifespan assay using 96-well liquid culture (100ul)?

Thanks.

2

I am not sure:

  1. Given that would have an additional mechano-sensory stress and I am not sure how worms react to additional force been applied on the worm cuticle and so on.
  2. Plus shaking them at this speeds would result in increase in temperature inside the wells.

Do you have any reference for the same?

3

I would try 150 or 200rpm at the most to give worms some agitation to float in the liquid medium.

4

There is at least one published protocol for doing lifespan assays in liquid culture in 96 well plates, and it doesn’t seem to require any shaking (except for a little bit of mixing when reagents are added). So regardless of whether it’s a good idea, you can apparently get on without it.

If you decide to go with shaking anyway - and it really might be a “good idea” - 900 rpm sounds like it might be excessive. If you have the time (and inclination), comparing results with and without shaking would of course be a good idea.

5

Thank you all for your suggestions !

6

do not shaking, using 24-well plate with 1mL medium works very well, better than 96well.