Flat agar and optimized media enable high-throughput C. elegans imaging in 96-well plates
Year: 2026
Type: other
Abstract: High-throughput imaging of Caenorhabditis elegans enables large-scale phenotypic screens of genetic mutations, drugs, and other perturbations — experiments that would be intractable to perform on 60 mm plates or slides. Many existing devices for high-throughput C. elegans imaging are incompatible with multichannel pipettes and liquid handlers, time-consuming to prepare, or susceptible to desiccation during long-term studies. A simpler alternative is to pour agar directly into 96-well plates. Unfortunately, agar forms a meniscus in the well, creating a curved surface that requires z-stack acquisition to capture all animals. We tested several ways to flatten the agar surface, including a mechanical press that we termed the “MeniSquish” and found that centrifuging agar in 8-well strips eliminated the meniscus and gave minimal tilt in the agar. This approach consistently yields full plates of flat wells suitable for single-plane imaging, using only equipment found in most labs and a 3D-printable accessory. During our work, we compiled several other ways to improve imaging conditions, such as reducing the concentration of phosphate and peptone in the media. Although we developed this for C. elegans, the protocol should benefit anyone imaging agar-dwelling organisms in high-throughput formats.
Source: Arcadia Science
Institution: Arcadia Science
Language: English
Subfield: Biophysics
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W7202118641
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