Tim Sanchez, Ph. D. , Emily A. Seidler, M. D. , David K. Gardner, D

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Presentation transcript:

Will noninvasive methods surpass invasive for assessing gametes and embryos? Tim Sanchez, Ph.D., Emily A. Seidler, M.D., David K. Gardner, D.Phil., Daniel Needleman, Ph.D., Denny Sakkas, Ph.D. Fertility and Sterility Volume 108, Issue 5, Pages 730-737 (November 2017) DOI: 10.1016/j.fertnstert.2017.10.004 Copyright © 2017 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Metabolic imaging on individual embryos. (A) FLIM measurements of nicotinamide adenine dinucleotide (NADH) or flavin adenine dinucleotide (FAD) produce fluorescence images, such as this NADH image of a mouse blastocyst. Because NADH and FAD are concentrated in the mitochondria, these images provide reflect mitochondrial distribution. Images can be segmented with processing techniques to identify the embryo and separate it from the background. (B) Photon arrival time counts that occurred within the embryo region can be pooled together into a single fluorescence-lifetime imaging microscopy histogram, representing the average state of the embryo. This histogram can be fit to a theoretical model of two exponentially decaying species–one for bound molecule and one for free. (C) From these fits, each measurement produces 4 quantitative parameters that are sensitive to changes in mitochondrial state. NADH and FAD measurements can also be taken in tandem for a total of up to 8 parameters. Bar = 20 μm. Fertility and Sterility 2017 108, 730-737DOI: (10.1016/j.fertnstert.2017.10.004) Copyright © 2017 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Example of digitalized metabolic imaging on individual 2-cell mouse embryos. (A) Fluorescence-lifetime imaging microscopy measurements of nicotinamide adenine dinucleotide and (B) flavin adenine dinucleotide produce fluorescence images that can subsequently be quantified by specific algorithms incorporating information related to the auto- fluorescent signal of each embryo (Fig. 1). Bar = 50 μm. Fertility and Sterility 2017 108, 730-737DOI: (10.1016/j.fertnstert.2017.10.004) Copyright © 2017 American Society for Reproductive Medicine Terms and Conditions