Purdue University, Department of Physics and Astronomy, West Lafayette, Indiana, United States.
Purdue University, Department of Animal Sciences, West Lafayette, Indiana, United States.
J Biomed Opt. 2019 Jun;24(6):1-4. doi: 10.1117/1.JBO.24.6.060502.
Early stage porcine parthenogenetic embryos were evaluated for metabolic activity using a biodynamic microscope (BDM) that images dynamic light scattering using low-coherence digital holography. The microscope has a 45-deg illumination configuration that reduces specular background for the imaging of small translucent samples. The off-axis illumination is compatible with coherence-gated imaging because of volumetric light scattering in which the coherence plane is tilted at half the illumination angle in a three-dimensional tissue target. The BDM was used to profile the viability of porcine parthenotes with normal and with inhibited mitochondrial adenosine triphosphate (ATP) production using Doppler fluctuation spectroscopy. The ATP concentrations in the parthenotes, which are indicative of developmental potential, were validated by a conventional bioluminescence assay. Biodynamic classifications achieved ∼80 % accuracy correlating sample ATP treatment, providing a quick, label-free surrogate measurement to replace invasive metabolic assays as a candidate for evaluating quality of early embryos in the assisted reproductive technology setting.
使用生物动力学显微镜（BDM）评估早期猪孤雌胚胎的代谢活性，该显微镜使用低相干数字全息术对动态光散射进行成像。该显微镜具有 45 度照明配置，可减少小半透明样品成像的镜面背景。离轴照明与相干门成像兼容，因为在三维组织目标中，相干平面以照明角度的一半倾斜，发生体积光散射。使用多普勒频移光谱法，BDM 用于分析具有正常和抑制线粒体三磷酸腺苷（ATP）产生的猪孤雌胚胎的活力。孤雌胚胎中的 ATP 浓度可通过传统的生物发光测定法进行验证，该浓度指示了胚胎的发育潜力。生物动力学分类的准确率达到约 80%，与样本 ATP 处理相关联，提供了一种快速、无标记的替代测量方法，可替代侵入性代谢测定法，作为评估辅助生殖技术中早期胚胎质量的候选方法。
