In vitro fertilization (IVF) and embryo transfer (ET) are well-established assisted reproductive techniques in the cattle industry

The protocols have matured through decades of improvements, yet the conception rate following IVF/ET in cattle is around 35-45%. The major limiting factors for ET and IVF are labor-intensive protocols, dependence on practitioner expertise, poor super ovulatory response, and inaccurate embryo assessment. This research effort aims to solve two of the significant challenges mentioned above by (1) improving the labor-intensive embryo dilution process using microfluidics and (2) developing a non-invasive and objective embryo assessment technique. Hawai'i innovation laboratory (HIL), in collaboration with agrimark genetics, whitaker embryo transfer services, and hawai'i pacific university, plans to develop a novel embryo assessment technique that is non-invasive and objective, unlike the subjective iets-approved morphological grading system. The proposed technique can assess the embryo viability quantifiable by measuring the electrical impedance and observation of morphokinetics such as blastocyst expansion rate, blastomere symmetry, and percentage of the intact embryonic cells represented by the extruded material in the perivitelline space. The embryo assessment takes place in a microfluidic device that can wash the embryo from the flushing medium, easing the embryo dilution process. Also, the microfluidic chip allows positioning a single bovine embryo between two micro-electrodes under optical microscopy, recording time-lapse images, analyzing morphokinetics data, and measuring electric impedance circumventing the background noise conductive media. In prior work, HIL has measured the electrical impedance of single artemia cysts at various phases of development in a microfluidic environment. A distinct pattern of impedance changes was observed at different stages of cyst development. The measured impedance changes corresponded to physiological changes as the cyst developed. The change in impedance during the first stage of development provided sufficient quantitative data to predict if the cyst would hatch. Also, in a recent work, the proposer experimented the morphokinetics observations through time-lapse image segmentation using deep neural network (DNN), which correlates with its genetic health. Building on the above research, HIL combines DNN-assisted morphokinetics and impedance spectroscopy and proposes developing a non-invasive, quantitative, embryo assessment technique that can determine viability by measuring internal cell information such as membrane capacitance cytoplasmic conductivity along with the morphokinetic data. If successful, this effort will be the path forward to a commercial tool for the ET/IVF practitioner and embryo producers. The tool will help determine bovine embryo viability quantitatively before the freezing step. Also, the tool can potentially improve the pregnancy rate by 10-15% per transfer by reducing misjudging and discarding of competent embryos and reducing transfer or less competent embryos. This work will help the nifa achieve its mission of fostering an innovative research strategy that may be used to develop safer and more effective means of regulating, preserving, or achieving animal reproduction.