High-throughput imaging flow cytometry by optofluidic time-stretch microscopy
High-throughput imaging flow cytometry by optofluidic time-stretch microscopy
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DOI：
10.1038/s41596-018-0008-7
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发表时间：
2018-07-01
期刊：
影响因子：
14.8
通讯作者：
Goda, Keisuke
中科院分区：
文献类型：
--
作者：
Lei, Cheng;Kobayashi, Hirofumi;Goda, Keisuke
在大的异质细胞群体中快速测定形态学和细胞内分子变异的能力对于理解和利用细胞异质性是必不可少的。光流体时间拉伸显微镜是实现这一目标的强大方法，因为它能够实现高通量成像流式细胞术，用于从人类血液到藻类的各种细胞类型的大规模单细胞分析，从而实现生物，医学，制药和绿色能源应用的独特类别。在这里，我们描述了如何通过光流体时间拉伸显微镜进行高通量成像流式细胞术。具体而言，该方案提供了关于如何构建光学时间拉伸显微镜和用于光流体时间拉伸显微镜的细胞聚焦微流控装置的分步说明，将其用于具有亚微米分辨率的高通量单细胞图像采集，每秒> 10，000个细胞，进行图像构建和增强，进行大规模单细胞分析的图像分析，并使用压缩感知和机器学习等计算工具来处理细胞“大数据”。假设所有组件都是现成的，一个由三到四名成员组成的研究团队，具有光学，电子，微流体，数字信号处理和样品制备的中级经验，可以在1个月的时间内完成该方案。
The ability to rapidly assay morphological and intracellular molecular variations within large heterogeneous populations of cells is essential for understanding and exploiting cellular heterogeneity. Optofluidic time-stretch microscopy is a powerful method for meeting this goal, as it enables high-throughput imaging flow cytometry for large-scale single-cell analysis of various cell types ranging from human blood to algae, enabling a unique class of biological, medical, pharmaceutical, and green energy applications. Here, we describe how to perform high-throughput imaging flow cytometry by optofluidic time-stretch microscopy. Specifically, this protocol provides step-by-step instructions on how to build an optical time-stretch microscope and a cell-focusing microfluidic device for optofluidic time-stretch microscopy, use it for high-throughput single-cell image acquisition with sub-micrometer resolution at >10,000 cells per s, conduct image construction and enhancement, perform image analysis for large-scale single-cell analysis, and use computational tools such as compressive sensing and machine learning for handling the cellular 'big data'. Assuming all components are readily available, a research team of three to four members with an intermediate level of experience with optics, electronics, microfluidics, digital signal processing, and sample preparation can complete this protocol in a time frame of 1 month.