PDMS filter structures for size-dependent larval sorting and on-chip egg extraction of C. elegans

Area of Science:

• Developmental Biology
• Microfluidics
• Biotechnology

Background:

• Standardized C. elegans assays necessitate age-synchronized populations to prevent age-induced phenotyping variability.
• Existing C. elegans synchronization methods are often labor-intensive, complex, and low-throughput.

Purpose of the Study:

• To develop a high-throughput, efficient, and rapid microfluidic device for sorting mixed larval stages of C. elegans.
• To enable user-friendly and reusable C. elegans population synchronization and embryo harvesting.

Main Methods:

• Fabrication of a polydimethylsiloxane (PDMS) microfluidic device using plasma activation and bonding.
• Utilizing passive hydrodynamic principles for sorting C. elegans larvae based on size and stage.
• Implementing an on-chip bleaching procedure for efficient embryo extraction and synchronized L1 larvae harvesting.

Main Results:

• The PDMS microfluidic device successfully sorted mixed larval populations of C. elegans into consecutive stages within minutes.
• The device allows for the collection of two distinct sorted larval populations from a single input.
• Cascading devices enabled efficient collection of target larvae from mixed populations, and rapid egg extraction yielded synchronized L1 larvae.

Conclusions:

• The developed microfluidic device offers a rapid, high-throughput, and user-friendly solution for C. elegans age synchronization.
• This technology significantly improves the efficiency and standardization of C. elegans-based research.
• The passive hydrodynamic sorting approach is reusable and adaptable for various C. elegans developmental stages.