Seeking Advice on Issues Encountered During C. elegans Lifespan Assays

Dear Professors and Researchers,

I am a novice in C. elegans cultivation and recently began conducting lifespan assays. In my experiments, I used FUdR-supplemented NGM solid media to effectively suppress egg-laying in the worms. However, I encountered a problem on days 4 and 5: the worms developed large protrusions near the gonad region, and their overall vitality seemed to deteriorate significantly.

I would like to ask if anyone has experienced similar issues under these conditions. Are there potential modifications to this method that could improve the outcomes? Additionally, I would greatly appreciate any suggestions or best practices for conducting lifespan assays with C. elegans.

Thank you in advance for your insights and guidance!

At which temperature do you keep the worms?

In our laboratory, we typically culture C. elegans at a temperature of 20°C

It might because of FUDR concentration or humility. Maybe you can decrease the concentration of FUDR (10um works well in our hand); or control the humility of your assay condition. Wet plates cause vulval protrusions problems. Hope it helps.

Oh! Thank you for your reply. The problem has been solved now. I found that adding FUdR too early would lead to a large number of Caenorhabditis elegans vulval explosions. Therefore, I adjusted the time of adding it to one day after the adult Caenorhabditis elegans started laying eggs, and now the phenomenon of vulval explosions has mostly disappeared.