PT  - JOURNAL ARTICLE
AU  - Ning Ma
AU  - Nabora Reyes de Mochel
AU  - Paula Duyen Anh Pham
AU  - Tae Yeon Yoo
AU  - Ken WY. Cho
AU  - Michelle A. Digman
TI  - Label-free assessment of pre-implantation embryo quality by the Fluorescence Lifetime Imaging Microscopy (FLIM)-phasor approach
AID  - 10.1101/286682
DP  - 2018 Jan 01
TA  - bioRxiv
PG  - 286682
4099  - http://biorxiv.org/content/early/2018/04/21/286682.short
4100  - http://biorxiv.org/content/early/2018/04/21/286682.full
AB  - 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.