Liang Fei, Zhu Junwen, Chai Huichao, Feng Yongxiang, Zhao Peng, Liu Shaofeng, Yang Yuanmu, Lin Linhan, Cao Liangcai, Wang Wenhui
State Key Laboratory of Precision Measurement Technology and Instrument, Department of Precision Instrument, Tsinghua University, Beijing, 100084, China.
Small Methods. 2023 Jul;7(7):e2201492. doi: 10.1002/smtd.202201492. Epub 2023 Mar 22.
Non-invasive and rapid imaging technique at subcellular resolution is significantly important for multiple biological applications such as cell fate study. Label-free refractive-index (RI)-based 3D tomographic imaging constitutes an excellent candidate for 3D imaging of cellular structures, but its full potential in long-term spatiotemporal cell fate observation is locked due to the lack of an efficient integrated system. Here, a long-term 3D RI imaging system incorporating a cutting-edge white light diffraction phase microscopy module with spatiotemporal stability, and an acoustofluidic device to roll and culture single cells in a customized live cell culture chamber is reported. Using this system, 3D RI imaging experiments are conducted for 250 cells and demonstrate efficient cell identification with high accuracy. Importantly, long-term and frequency-on-demand 3D RI imaging of K562 and MCF-7 cancer cells reveal different characteristics during normal cell growth, drug-induced cell apoptosis, and necrosis of drug-treated cells. Overall, it is believed that the proposed 3D tomographic imaging technique opens up a new avenue for visualizing intracellular structures and will find many applications such as disease diagnosis and nanomedicine.
在亚细胞分辨率下的非侵入性快速成像技术对于细胞命运研究等多种生物学应用具有重要意义。基于无标记折射率（RI）的三维断层成像构成了细胞结构三维成像的优秀候选方法，但由于缺乏高效的集成系统，其在长期时空细胞命运观察中的全部潜力尚未得到发挥。在此，报道了一种长期三维RI成像系统，该系统结合了具有时空稳定性的前沿白光衍射相显微镜模块以及用于在定制的活细胞培养室中滚动和培养单细胞的声流体装置。使用该系统，对250个细胞进行了三维RI成像实验，并展示了高效且高精度的细胞识别。重要的是，对K562和MCF - 7癌细胞的长期按需频率三维RI成像揭示了正常细胞生长、药物诱导的细胞凋亡以及药物处理细胞坏死过程中的不同特征。总体而言，人们认为所提出的三维断层成像技术为可视化细胞内结构开辟了一条新途径，并将在疾病诊断和纳米医学等诸多领域得到应用。
