Improving IVF Success through Multiplexed Optical Metabolic Imaging - Summary In assisted reproductive technology (ART), morphological evaluation by bright- or dark-field optical microscopy is the most critical assessment on the implantation potential (viability) of an in vitro fertilized embryo. Ideally the obtained morphological information should be complemented with spatially resolved metabolic or functional information. This would provide a more complete picture of the embryo itself and may thus offer a more objective viability assay to increase the successful rate of an embryo transfer toward (singleton) live-birth. Currently, fertility clinic providers are unable to visualize embryonic metabolism, whereas the spectroscopy- based metabolic profiling under development has lacked spatial resolution. Therefore, a critical need exists to superimpose spatially resolved metabolic information on morphological information for full assessment of the embryo before implantation. An interdisciplinary partnership composed of collaborating investigators in optical instrumentation, reproductive biology, and computer science will address this critical need by multiplexed optical metabolic imaging (MOMI). In contrast to the intrinsically label-free bright- or dark-field optical microscopy unable to obtain molecular information and resolve a three–dimensional structure, MOMI enables these abilities via well-known endogenous metabolites and nonlinear optical excitation, respectively, while critically retains the label-free aspect required for potential clinical application. The preliminary results have demonstrated the utility of MOMI as a general imaging assay for unlabeled live cells and cultured spheroids or organoids. This partnership will thus aim to validate the MOMI in an established bovine model of ART. Future plans are to extend these methodologies to improve human ART. The goal of this partnership will be achieved through a systematic approach. First, the inclusion of fluorescence lifetime imaging will upgrade MOMI to an omics-like assay, and a dedicated environment of incubation will be built to interface the imaging with preimplantation embryo culturing. Also, an established bovine model for human ART will be employed to image 2500 preimplantation embryos and track subsequent live births of ~1000 embryo transfers. Finally, a novel machine learning algorithm based on multi-cell graph neural network from MOMI images will be developed to predict the viability of a preimplantation embryo (prospectively). The successful completion of this project will validate MOMI as a safe live-cell imaging assay for non-ART biomedical applications such as cell therapy and drug screening, considering the embryo development as a well-known sensitive bioassay for plausible optical imaging-induced phototoxicity. More importantly, this large- scale preclinical study will justify a randomized clinical trial for human ART. Ultimately, the clinical application of MOMI may significantly improve the success rate of live births versus embryo transfers that has remained stagnant in the last decade (35-37%) and may therefore reduce the associated pain and cost.
