PROJECT SUMMARY Objective Biotechnology is a spin-out from the University of Minnesota that is pioneering the use of machine vision guided robots for automating precise microbiology procedures such as microinjection. Drosophila melanogaster (fruit fly) is a model organism extensively used in both basic and clinical research. A key challenge in Drosophila biological research is the bottleneck of generating and maintaining transgenic lines of flies. Traditionally, transgenesis involves skilled technicians performing intricate and precise microinjection procedures repeatedly. However, Objective Biotechnology Inc. has introduced an innovative solution - a machine vision guided robot designed to automate the microinjection process for Drosophila melanogaster embryos. This technology can also be adapted for use with various other organisms. This robot eliminates the need for manual microinjection protocols, which are operator-dependent, time-consuming, and require significant training. The robot uses machine learning (ML) models trained to detect individual embryos on agar plates and guides microinjection needles to perform microinjections at specific locations in each detected embryo. This robot can be operated by individuals with no prior experience and surpass human capabilities in terms of microinjection speed, performing at a rate six times faster than humans. In AIM 1 of this proposal, we will evaluate the efficacy and generalizability of the Autoinjector technology for transgenesis across a wide spectrum of Drosophila experiments. This involves testing and refining the automated microinjection process for different genetic backgrounds, microinjection locations, microinjectant compositions, construct sizes, auxiliary plasmids or transgenes, as well as various operational variables such as embryo laying conditions, culture media, DNA concentration, and solution viscosity. AIM 2 of this proposal we will innovate the ML algorithms to address two identified failure modes in the automated microinjection process. These failure modes are (1) the inability to detect individual embryos when they are clustered together on the embryo collection plates and (2) the clogging of pipettes during automated microinjection. We will develop ML-guided robotic algorithms to mitigate these issues and improve the overall performance of the system.

PROJECT NARRATIVE In this proposal, Objective Biotechnology seeks to develop a commercial-grade automated microinjection system, building on an extensively validated system developed by the company’s founders at the University of Minnesota via an R21 grant. This technology will support microinjection investigations for transgenesis, mutagenesis, cell labeling, small molecule screening, cryopreservation, and in vitro fertilization across many different organisms. Our automated microinjection system will make these techniques more accessible, reduce costs, and dramatically increase the scale at which such experiments can be carried out, broadly accelerating discovery related to human health and disease in animal research models, and thus this proposal is relevant to most NIH Institutes.