High-throughput genetic manipulation of multicellular organisms using a machine-vision guided embryonic...

Area of Science:

• Biotechnology and Biomedical Engineering
• Developmental Biology
• Genetics

Background:

• Microinjection is crucial for transgenesis, mutagenesis, and cryopreservation but requires specialized skills and is labor-intensive.
• Current microinjection methods are rate-limiting and pose challenges for large-scale experiments.

Purpose of the Study:

• To develop and validate a machine-vision guided robotic system for fully automated microinjection in Drosophila melanogaster and Danio rerio embryos.
• To enhance throughput, efficiency, and success rates of microinjection for transgenesis and cryopreservation.

Main Methods:

• Construction of a generalized robot utilizing machine learning for embryo detection and 3D anatomical localization.
• Dual-view microscopy and automated serial microinjection of tens of thousands of embryos.
• Optimization of robotic microinjection protocols for fruit flies and zebrafish.

Main Results:

• Robotic microinjection achieved throughput increases of up to 4× in Drosophila compared to manual methods.
• Generation of over 400 unique transgenic Drosophila lines from 1,713 injected embryos using barcoded plasmids.
• Significantly improved vitrification rates and post-thaw survival for cryopreserved zebrafish embryos via robotic microinjection.

Conclusions:

• The developed robotic system automates microinjection with proficiency comparable to human experts, enabling large-scale genetic manipulation and cryopreservation.
• This technology offers a scalable solution for genome-wide studies and efficient cryopreservation across various organisms.
• Robotic microinjection advances research capabilities in developmental biology and genetic engineering.