Category: Assay Development and Screening
Conventional techniques for femtosecond laser axotomy in the nematode C. elegans require manual orientation and immobilization of the animals for surgery and post-recovery imaging. To overcome these time-consuming and labor-intensive processes, several microfluidic methods have become available, albeit with multi-layered designs that are often too complicated to manufacture and operate for non-expert users. Here we present a simpler device, consisting of a single polydimethylsiloxane (PDMS) layer, which can immobilize up to 20 animals. The animals are trapped in the favorable orientation for optical access needed for precise laser surgery and high-resolution imaging. The new device, named “worm hospital” allows on-chip axotomy, post-surgery housing for recovery, and post-recovery imaging of nerve regeneration all on one microfluidic chip. Utilizing the worm hospital and analysis of mutants, we observed that most but not all neurodevelopmental genes in the Wnt/Frizzled pathway are important for regeneration of the two touch receptor neurons ALM and PLM. Using our new chip, we observed that the cwn-2 and cfz-2 mutations significantly reduced the reconnection possibilities of both neurons without any significant reduction in the regrowth lengths of the severed axons. We observed a similar regeneration phenotype with cwn-1 mutation in ALM neurons only. Our platform provides an opportunity to screen C. elegans models for novel drug candidates and genetic interactors using RNA interference in a high-content manner.
Sudip Mondal– Research Associate, UT Austin, Austin, TX
I obtained a Ph.D. from IISc, India in Physics in the year 2008 and joined NCBS-TIFR, India for a postdoctoral research to gain insight in C. elegans neurobiology and developed a microfluidic platform to study in vivo neuronal transport of various organelles. Currently I am a research associate at UT Austin where I am developing large-scale microfluidic platforms for high-throughput drug screening using C. elegans as a disease model. Specifically, we plan to identify new chemical compounds and genetic players with protection against various age-dependent degeneration model and regeneration in C. elegans. We are also interested in developing multiple imaging modalities for rapid screening of C. elegans model for deep phenotyping. I have developed a wide range of research experience in the field of microfluidics technology, imaging, C. elegans neurobiology, and high-throughput assay development etc. My research work enables large-scale high-content screening of C. elegans disease models.