For this assay we take advantage of the existing C. elegans GFP-reporter strains, modelling a variety of neurodegenerative disorders, such as Alzheimer’s, Parkinson’s, Huntington’s disease and amyotrophic lateral sclerosis (ALS), as well as the capacity of our device to perform fluorescent imaging.
A synchronized population of C. elegans is injected into the microfluidic platform at the first larval stage (L1). Worms are then confined within dedicated microfluidic chambers and are continuously fed with an E. coli solution.
Worms are chronically exposed to the test compounds starting from the last larval stage prior to sexual maturity (L4) for 85 hours (day 3 of adulthood). The compounds to be tested are mixed with the E. coli solution. Freeze-dried OP50 E. coli are used as a food source for the whole duration of the experiment, preventing the metabolization of the tested molecules by the bacteria.
The images of each microfluidic chamber are recorded every hour. Starting from 70 hours post worm injection (day 1 of adulthood), both brightfield and fluorescent images start to be recorded in order to monitor worm growth and protein aggregation dynamics in parallel. Time-resolved phenotypic readouts are then extracted from the collected images in both brightfield and fluorescent modes.
- Aggregate number
- Aggregate size