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bioRxiv · 10.1101/286682

Label-free assessment of pre-implantation embryo quality by the Fluorescence Lifetime Imaging Microscopy (FLIM)-phasor approach

Ma, N.·Reyes de Mochel, N.·Pham, P.·Yoo, T.·Cho, K.·Digman, M.
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Abstract

Development of quantitative, safe and rapid techniques for assessing embryo quality provides significant advances in Assisted Reproductive Technologies (ART). We apply the phasor-FLIM method to capture endogenous fluorescent biomarkers of pre-implantation embryos as a non-morphological caliber for embryo quality. Here, we identify the developmental, or \"D-trajectory\", that consists of fluorescence lifetime from different stages of mouse pre-implantation embryos. The D-trajectory correlates with intrinsic fluorescent species from a distinctive energy metabolism and oxidized lipids, as seen with Third Harmonic Generation (THG) that changes over time. In addition, we have defined an Embryo Viability Index (EVI) to distinguish pre-implantation embryo quality using the Distance Analysis, a machine learning algorithm to process the fluorescence lifetime distribution patterns. We show that the phasor-FLIM approach provides a much-needed non-invasive quantitative technology for identifying healthy embryos at the early compaction stage with 86% accuracy. This may increase embryo implantation success for in vitro fertilization clinics.\n\nHighlightsO_LIA label-free method of tracking metabolic trajectories during pre-implantation mouse embryo development.\nC_LIO_LIA non-invasive approach for assessing embryo quality and viability by a phasor-FLIM analysis.\nC_LI

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Ma, N., Reyes de Mochel, N., Pham, P., Yoo, T., Cho, K., Digman, M.. 2018-03-22. Label-free assessment of pre-implantation embryo quality by the Fluorescence Lifetime Imaging Microscopy (FLIM)-phasor approach. https://doi.org/10.1101/286682

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