Beckman Institute for Advanced Science and Technology, The University of Illinois at Urbana-Champaign, Urbana, IL 61801.
Department of Electrical and Computer Engineering, The University of Illinois at Urbana-Champaign, Urbana, IL 61801.
Proc Natl Acad Sci U S A. 2020 Aug 4;117(31):18302-18309. doi: 10.1073/pnas.2001754117. Epub 2020 Jul 20.
The ability to evaluate sperm at the microscopic level, at high-throughput, would be useful for assisted reproductive technologies (ARTs), as it can allow specific selection of sperm cells for in vitro fertilization (IVF). The tradeoff between intrinsic imaging and external contrast agents is particularly acute in reproductive medicine. The use of fluorescence labels has enabled new cell-sorting strategies and given new insights into developmental biology. Nevertheless, using extrinsic contrast agents is often too invasive for routine clinical operation. Raising questions about cell viability, especially for single-cell selection, clinicians prefer intrinsic contrast in the form of phase-contrast, differential-interference contrast, or Hoffman modulation contrast. While such instruments are nondestructive, the resulting image suffers from a lack of specificity. In this work, we provide a template to circumvent the tradeoff between cell viability and specificity by combining high-sensitivity phase imaging with deep learning. In order to introduce specificity to label-free images, we trained a deep-convolutional neural network to perform semantic segmentation on quantitative phase maps. This approach, a form of phase imaging with computational specificity (PICS), allowed us to efficiently analyze thousands of sperm cells and identify correlations between dry-mass content and artificial-reproduction outcomes. Specifically, we found that the dry-mass content ratios between the head, midpiece, and tail of the cells can predict the percentages of success for zygote cleavage and embryo blastocyst formation.
在高通量水平上评估精子的能力将对辅助生殖技术(ART)非常有用,因为它可以允许对体外受精(IVF)进行特定的精子选择。在生殖医学中,内在成像和外部对比剂之间的权衡尤为突出。荧光标记的使用使新的细胞分选策略成为可能,并为发育生物学提供了新的见解。然而,对于常规临床操作,使用外在对比剂通常过于侵入性。这引发了关于细胞活力的问题,尤其是对于单细胞选择,临床医生更喜欢以相差、微分干涉对比或霍夫曼调制对比形式存在的内在对比。虽然这些仪器是非破坏性的,但得到的图像缺乏特异性。在这项工作中,我们提供了一个模板,通过将高灵敏度相位成像与深度学习相结合,来规避细胞活力和特异性之间的权衡。为了为无标记图像引入特异性,我们训练了一个深度卷积神经网络在定量相位图上执行语义分割。这种方法,一种具有计算特异性的相位成像(PICS),使我们能够有效地分析数千个精子细胞,并确定干物质含量与人工繁殖结果之间的相关性。具体来说,我们发现细胞头部、中段和尾部之间的干物质含量比值可以预测受精卵分裂和胚胎囊胚形成的成功率百分比。