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

Genetics, Journal Year: 2024, Volume and Issue: 226(4)

Published: Feb. 19, 2024

Abstract Microinjection is a technique used for transgenesis, mutagenesis, cell labeling, cryopreservation, and in vitro fertilization multiple single multicellular organisms. requires specialized skills involves rate-limiting labor-intensive preparatory steps. Here, we constructed machine-vision guided generalized robot that fully automates the process of microinjection fruit fly (Drosophila melanogaster) zebrafish (Danio rerio) embryos. The uses machine learning models trained to detect embryos images agar plates identify specific anatomical locations within each embryo 3D space using dual view microscopes. then serially performs detected embryo. We three such robots automatically microinject tens thousands Drosophila systematically optimized robotic species performed routine transgenesis with proficiency comparable highly skilled human practitioners while achieving up 4× increases throughput Drosophila. was utilized pools over 20,000 uniquely barcoded plasmids into 1,713 2 days rapidly generate more than 400 unique transgenic lines. This experiment enabled novel measurement number independent germline integration events per successfully injected Finally, showed cryoprotective agents significantly improves vitrification rates survival cryopreserved post-thaw as compared manual microinjection. anticipate can be carry out genome-wide manipulation cryopreservation at scale wide range

Language: Английский

A machine vision guided robot for fully automated embryonic microinjection

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: April 25, 2023

ABSTRACT Microinjection is a widely used technique for transgenesis, mutagenesis, cell labeling, cryopreservation, and in vitro fertilization multiple single multicellular organisms. requires specialized skills acquired each target organism involves rate limiting labor-intensive preparatory steps. Here we constructed machine vision (MV) guided generalized robot that fully automates the process of microinjection fruit fly (Drosophila melanogaster) zebrafish ( Danio rerio ) embryos. The uses learning (ML) models trained to detect individual embryos images agar plates, identify specific anatomical locations within embryo 3D space using dual view microscopes. this information serially perform detected without any human intervention. We three such robots automatically microinject tens thousands Drosophila systematically optimized robotic species validated use by performing routine transgenesis with proficiency comparable highly skilled practitioners while achieving up 4x increases throughput Drosophila. automated was utilized pools over 20,000 uniquely barcoded plasmids into 1,713 two days rapidly generate more than 400 unique transgenic lines. This high experiment enabled novel measurement number independent germline integration events per successfully injected embryo. Finally, showed cryoprotective agents significantly improves vitrification rates survival cryopreserved post-thaw as compared manual microinjection, opening tantalizing possibility large-scale cryobanking aquatic at an industrial scale. anticipate versatile system can be applied carry out genome-wide manipulation cryopreservation scale wide range other

Language: Английский