Counting the Dead and Living worms in an Anthelmintic Assay

I am working on an Anthelmintic assay for which I am using C.elegans as a model organism. The positive control I am using is Mebendazole. Recently I tested the drug on 10 worms. After an incubation period of around 5 days, I found that 6 to 7 of them died. But around 4 to 5 of them died even in the Negative control and the plate without an intervention. I did not use any OP50 as the feed. The worms were suspended in M9 solution. It was a very rough experiment conducted just to see how the Anthelmintic activity really works. The concentration of Mebendazole I used was around 5 Micrograms per 1ml of DMSO. I am not very convinced with the experiment results. Shall I increase the concentration of Mebendazole i am using? I am not sure if the worms died because of the Mebendazole or because of starvation. Anyway, now i will be conducting the same experiment on nearly 500 worms. Here counting the dead and live worms is an issue. Is there any easier method to quantify the number of dead and living worms? apart from visual counting of course… does anyone have any Information about this???

You might look for ideas in this previous thread

Any direct method that can avoid the manual counting of the dead and live worms and make the process comparatively less laborious?
Could not find satisfactory ideas in the suggested thread…


If you can do it in a liquid assay, sytox green at 10um will cause dead worms to fluoresce and does not affect the live worms.