论文标题
拉曼 - 大气流体集成系统用于单细胞分析
Raman-acoustofluidic integrated system for single-cell analysis
论文作者
论文摘要
共聚焦拉曼显微镜提供了一种特定的途径,用于监测细胞膜,细胞器和细胞核等细胞内成分的化学“指纹”。然而,用于生物医学研究的固定或随机分散细胞(例如显微镜载玻片)上的固定或随机分散细胞(例如显微镜载玻片)的常规拉曼采集可能受到严重限制。反向散射的拉曼信号可能会被底物荧光的贡献所掩盖。同样,用于药物发现,感染和组织工程的生物测定可能需要在几个小时到几天的时间间隔内单独监测活细胞。为了满足细胞测定监测的需求,我们提出了一种大声液体装置,该设备在10μl体积的圆柱室中形成悬浮的细胞聚集(一层),该腔室以1 MHz的频率工作。集成的系统包括一个实验室芯片设备和共聚焦拉曼显微镜。在此设置中,可以在没有任何底物干扰的情况下以微米准确性来选择性地对Raman进行访问。基于一组精心设计的实验,我们证明了聚苯乙烯微粒被组装并保持停滞,使得能够在不到一分钟的时间内从单个粒子中摄取完整的拉曼光谱。与传统的拉曼采集相比,信噪比的比率提高了千倍。展示了Raman-Acoustofluidic系统,用于在富集的小鼠巨噬细胞中获得单个细胞的光谱。获得的结果证实了该方法在单细胞分析中应用的鲁棒性。
Confocal Raman microscopy offers a particular pathway for monitoring chemical "fingerprints" of intracellular components like the cell membrane, organelles, and nucleus. Nevertheless, conventional Raman acquisitions of fixed or randomly dispersed cells on a substrate, such as a microscope slide, might be severely limited for biomedical investigations. Backscattered Raman signal is likely to be overshadowed by contributions from substrate fluorescence. Also, biological assays for drug discovery, infection, and tissue engineering may require monitoring live cells individually over a time interval spanning from a few hours to a few days. To meet the needs of cell assay monitoring, we propose an acoustofluidic device that forms a levitating cell aggregation (one layer) in a cylindrical chamber of 10 μL volume that operates at 1 MHz frequency. The integrated system comprises a lab-on-a-chip device and a confocal Raman microscope. In this setup, a cell can be selectively Raman-investigated with micrometre accuracy, without any substrate interference. Based on a set of carefully designed experiments, we demonstrate that polystyrene microparticles are assembled and held standstill, enabling a full Raman spectrum to be taken of a single particle in less than a minute. The signal-to-background ratio is improved by a thousandfold when compared with conventional Raman acquisition. The Raman-acoustofluidic system is showcased for obtaining the spectrum of a single cell among an enriched population of macrophages of mice. The obtained results confirm the method's robustness for applications in single-cell analysis.