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A Semi-high-throughput Imaging Method and Data Visualization Toolkit to Analyze C. elegans Embryonic Development
Published on: October 29, 2019
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High-Temporal-Resolution smFISH Method for Gene Expression Studies in Caenorhabditis elegans Embryos
Seleipiri Charles1,2, Guillaume Aubry3, Han-Ting Chou4
1Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, 313 Ferst Drive NW, Atlanta, Georgia 30332, United States.
Analytical Chemistry
|December 23, 2020
Summary
This study introduces a microfluidic pipeline for high-throughput single-molecule fluorescence in situ hybridization (smFISH) in Caenorhabditis elegans embryos, significantly reducing labor and increasing experimental scale for developmental studies.
Area of Science:
- Developmental Biology
- Molecular Biology
- Bioengineering
Background:
- Fluorescence-based molecular tools are crucial for developmental studies but often require labor-intensive sample preparation.
- Large sample sizes are essential for detecting subtle phenotypes and gene expression dynamics in embryogenesis.
- Current methods, like single-molecule fluorescence in situ hybridization (smFISH) in C. elegans, face challenges with scalability due to manual sample handling.
Purpose of the Study:
- To develop a microfluidic pipeline for large-scale, labor-minimized smFISH imaging of C. elegans embryos.
- To overcome the bottleneck of efficient reagent exchange for numerous samples in microfluidic devices.
- To enhance throughput and enable high-temporal-resolution longitudinal studies in developmental biology.
Main Methods:
- Designed a microfluidic device with embryo traps for simultaneous processing of hundreds of C. elegans embryos.
- Engineered a protocol for efficient chemical exchange within the microfluidic device.
- Validated the preservation of image quality and demonstrated increased throughput compared to manual methods.
Main Results:
- The microfluidic pipeline enables large-scale smFISH imaging with minimized user input and labor.
- Efficient reagent exchange was achieved for hundreds of embryos simultaneously on-chip.
- The device significantly increases throughput, replacing the equivalent of 10 glass slides per experiment.
- Image quality was preserved, and the platform demonstrated capability for longitudinal gene expression analysis (e.g., par-1).
Conclusions:
- The developed microfluidic approach substantially enhances the throughput of smFISH in C. elegans embryogenesis.
- This method facilitates systematic, high-temporal-resolution studies, benefiting large-scale genetic screens and drug discovery.
- The platform is adaptable for various biological systems beyond C. elegans embryos.
