具有光散射参考的光声流式细胞术
Optoacoustic flow-cytometry with light scattering referencing
摘要：
与荧光蛋白的发展类似，用于筛选光声细胞标记的创新工具可以为OA提供定制的蛋白质标记，从而提供可视化生物结构和功能的新方法。光声成像以其优越的性能，如可达深度和活组织的3D研究，成为生命科学和医疗实践中一种非常有前途的模式。开发具有分子特异性的新型标记物可以显著增强光声对比度、特异性和灵敏度，并允许光声询问不适合荧光方法的组织。我们报告的光声流式细胞仪（OAFC）的原型，筛选光声报告基因。细胞计数器同时记录光散射以用于参考目的。由于记录光散射完全独立于OA，我们相信它是比例如荧光或超声反向散射更可靠的参考方法。我们的OAFC原型的精确表征展示了其光声表征单细胞大小范围内的流动物体的能力。我们应用OAFC来区分个体E。大肠杆菌细胞基于其表达的色蛋白的光声特性，使用微流体装置读取流中，并实现了90%以上的精度。我们讨论了如何光散射参考OAFC方法提供了一个关键的步骤，对常规测量的单细胞的光声特性，并可能铺平道路，识别基因编码的光声记者，通过转移荧光场的工作概念。
英文摘要：
In analogy to the development of fluorescent proteins, innovative tools for screening optoacoustic cell labels could lead to tailored protein labels for OA, imparting novel ways to visualize biological structure and function. Optoacoustic imaging emerges towards a highly promising modality for life sciences and medical practise with advantageous capabilities such as great accessible depth, and 3D studying of living tissue. The development of novel labels with molecular specificity could significantly enhance the optoacoustic contrast, specificity, and sensitivity and allow optoacoustic to interrogate tissues not amenable to the fluorescence method. We report on an optoacoustic flow cytometer (OAFC) prototype, developed for screening optoacoustic reporter genes. The cytometer concurrently records light scattering for referencing purposes. Since recording light scattering is completely independent from OA, we believe it to be a more reliable referencing method than e.g. fluorescence or ultrasound-backscatter. Precise characterization of our OAFC prototype showcases its ability to optoacoustically characterize objects in-flow that are in the size range of single cells. We apply the OAFC to distinguish individual E. coli cells based on optoacoustic properties of their expressed chromoproteins read in-flow using microfluidic arrangements and achieved precisions over 90%. We discuss how the light scattering referenced OAFC method offers a critical step towards routine measurement of optoacoustic properties of single-cells and could pave the way for identifying genetically encoded optoacoustic reporters, by transferring working concepts of the fluorescence field.
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