LED based real-time survival bioassays for nematode research

Nematode bioassays are extensively conducted worldwide, either for screening anthelmintic drugs or for assessing the toxicity of drug candidates. Recently, the US Environmental Protection Agency mandated the use of invertebrate models including nematodes especially Caenorhabditis elegans, for toxicity testing as an alternative to rodent models. The significance of nematode bioassays in the biological sciences is escalating, but no standardized protocol is available to assess nematode mortality in a liquid medium. Manual counting under white light is the only approach currently practiced, which exhibit large variabilities and false positive results. Here, we describe an innovative counting strategy that employs light-emitting diode (LED) technology. We found that the nematodes stopped moving under white light (360–760 nm) when administered with sub-lethal dosage (LC(50)) of a toxic drug, whereas they responded rapidly to blue (450–490 nm) and ultraviolet (UV) (100–400 nm) LED lights. Furthermore, paralyzed nematodes responded in less than 5 seconds to a LED pulse. The response to the LED stimulus was distinctively noted in C. elegans dauers, which squirmed away from illuminated sites within seconds. LED produced an incoherent beam, and uniformly distributed light across the sampling area. In conclusion, this method is more accurate than the conventional counting techniques, and enables us to differentiate paralyzed and dead nematodes virtually in real-time. Furthermore, this technique would appear to be suitable for incorporating a motion-sensor based automated system.