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

Ning Ma, Nabora Reyes de Mochel, Paula Duyen Anh Pham, Tae Yeon Yoo, Ken WY. Cho, Michelle A. Digman
doi: https://doi.org/10.1101/286682
Ning Ma
1Department of Biomedical Engineering, University of California, Irvine, CA
2Laboratory of Fluorescence Dynamics (LFD)
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Nabora Reyes de Mochel
3Department of Developmental and Cell Biology, University of California, Irvine, CA
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Paula Duyen Anh Pham
3Department of Developmental and Cell Biology, University of California, Irvine, CA
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Tae Yeon Yoo
3Department of Developmental and Cell Biology, University of California, Irvine, CA
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Ken WY. Cho
3Department of Developmental and Cell Biology, University of California, Irvine, CA
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  • For correspondence: mdigman@uci.edu kwcho@uci.edu
Michelle A. Digman
1Department of Biomedical Engineering, University of California, Irvine, CA
2Laboratory of Fluorescence Dynamics (LFD)
3Department of Developmental and Cell Biology, University of California, Irvine, CA
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  • For correspondence: mdigman@uci.edu kwcho@uci.edu
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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.

  • A label-free method of tracking metabolic trajectories during pre-implantation mouse embryo development.

  • A non-invasive approach for assessing embryo quality and viability by a phasor-FLIM analysis.

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Posted April 21, 2018.
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Label-free assessment of pre-implantation embryo quality by the Fluorescence Lifetime Imaging Microscopy (FLIM)-phasor approach
Ning Ma, Nabora Reyes de Mochel, Paula Duyen Anh Pham, Tae Yeon Yoo, Ken WY. Cho, Michelle A. Digman
bioRxiv 286682; doi: https://doi.org/10.1101/286682
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Label-free assessment of pre-implantation embryo quality by the Fluorescence Lifetime Imaging Microscopy (FLIM)-phasor approach
Ning Ma, Nabora Reyes de Mochel, Paula Duyen Anh Pham, Tae Yeon Yoo, Ken WY. Cho, Michelle A. Digman
bioRxiv 286682; doi: https://doi.org/10.1101/286682

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