Femtosecond laser microdissection isolates single regenerating C. elegans neurons for single cell RNA-sequencing

This project aims to establish femtosecond laser microdissection (fs-LM) as a new method that can isolate intact cells directly from living tissue. C. elegans has been established as a versatile model for nerve regeneration studies. Due to a lack of method, it has been infeasible to isolate regenerating C. elegans neurons for molecular profiling.We demonstrated feasibility and advantage of fs-LM by comparing gene expression in single C. elegans touch receptor neurons isolated by fs-LM and the dissociation-FACS method. In addition, we used fs-LM to isolate both uninjured and laser axotomized PLM neurons from wild-type and DLK-1 knockout animals to reveal transcriptional activities underlying nerve regeneration in C. elegans.