Label-free chemical imaging flow cytometry by high-speed multicolor stimulated Raman scattering
Label-free chemical imaging flow cytometry by high-speed multicolor stimulated Raman scattering
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DOI：
10.1073/pnas.1902322116
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发表时间：
2019-08-06
影响因子：
11.1
通讯作者：
Ozeki, Yasuyuki
中科院分区：
文献类型：
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作者：
Suzuki, Yuta;Kobayashi, Koya;Ozeki, Yasuyuki
成像流式细胞术将流式细胞术与荧光成像和数字图像分析的优势相结合，是癌症生物学、免疫学、药物发现、微生物学和代谢工程等多个领域的强大工具。它能够对众多活细胞的化学、结构和形态表型进行测量和统计分析，从而提供对生物过程的系统见解。然而，它的实用性受到表型分析荧光标记要求的限制。在这里，我们提出无标记化学成像流式细胞术来克服这个问题。它建立在脉冲对分辨波长可切换斯托克斯激光器的基础上，可在 3D 声学聚焦微流体芯片上对快速流动的细胞进行迄今为止最快的多色受激拉曼散射 (SRS) 显微镜检查，从而实现高达 140 个细胞/秒的前所未有的吞吐量。为了展示其广泛的实用性，我们使用 SRS 成像流式细胞术并借助深度学习来研究微藻细胞的代谢异质性并在血液中进行无标记癌症检测。
Combining the strength of flow cytometry with fluorescence imaging and digital image analysis, imaging flow cytometry is a powerful tool in diverse fields including cancer biology, immunology, drug discovery, microbiology, and metabolic engineering. It enables measurements and statistical analyses of chemical, structural, and morphological phenotypes of numerous living cells to provide systematic insights into biological processes. However, its utility is constrained by its requirement of fluorescent labeling for phenotyping. Here we present label-free chemical imaging flow cytometry to overcome the issue. It builds on a pulse pair-resolved wavelength-switchable Stokes laser for the fastest-to-date multicolor stimulated Raman scattering (SRS) microscopy of fast flowing cells on a 3D acoustic focusing microfluidic chip, enabling an unprecedented throughput of up to similar to 140 cells/s. To show its broad utility, we use the SRS imaging flow cytometry with the aid of deep learning to study the metabolic heterogeneity of microalgal cells and perform marker-free cancer detection in blood.