Spatial light-modulated stimulated Raman scattering (SLM-SRS) microscopy for rapid multiplexed vibrational imaging
Spatial light-modulated stimulated Raman scattering (SLM-SRS) microscopy for rapid multiplexed vibrational imaging
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DOI：
10.7150/thno.38551
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发表时间：
2020-01-01
期刊：
影响因子：
12.4
通讯作者：
Huang, Zhiwei
中科院分区：
文献类型：
--
作者：
Bae, Kideog;Zheng, Wei;Huang, Zhiwei
高速成像是监测生物事件中动态过程的先决条件。在这里，我们报告了一种独特的空间光调制受激拉曼散射（SLM-SRS）显微镜的发展，该显微镜将宽带激发光束与设计用于快速多路复用振动成像的稀疏采样掩模相结合，以监测实时癌症治疗效果和药物溶剂的体内运输。我们设计了一个最佳的掩模图案，使得能够选择SRS光谱中的主要窗口，以最高可能的峰值功率进行集体激发，从而提供了改善的信噪比（SNR）而不损害化学特异性。使用柔性空间光调制器将所生成的掩模图案应用于宽激发光束。SLM模块还提供了补充功能，从而可以促进快速扫描的SRS光谱的掩模生成之前，从而使SLM-SRS系统的一个独立的imaging platform.Results：我们表明，SLM-SRS显微镜允许快速复用的SRS成像聚苯乙烯和聚甲基丙烯酸甲酯珠在布朗运动在二甲基亚砜（DMSO）在70毫秒的时间间隔没有运动artifacts。我们进一步应用SLM-SRS监测弱碱性溶液对癌细胞的治疗作用，显示出即时的凋亡反应。最后，我们可视化在体内渗透到植物组织中的DMSO和评估急性毒性的DMSO对纤维素和蛋白质内tissue.Conclusion：我们开发了新的SLM-SRS显微镜和确认其广泛的适用性，在亚细胞和分子水平上的动态生物过程的快速监测。
High speed imaging is pre-requisite for monitoring of dynamic processes in biological events. Here we report the development of a unique spatial light-modulated stimulated Raman scattering (SLM-SRS) microscopy that tailors the broadband excitation beam with sparse-sampling masks designed for rapid multiplexed vibrational imaging to monitor real-time cancer treatment effects and in vivo transport of drug solvent.Methods: We design an optimal mask pattern that enables selection of predominant windows in SRS spectrum for collective excitation at the highest possible peak power, thus providing an improved signal-to-noise ratio (SNR) without compromise of chemical specificity. The mask pattern generated is applied to the broad excitation beam using a flexible spatial light modulator. The SLM module further offers complementary function whereby rapid scanning of SRS spectrum can be facilitated prior to the mask generation, thereby making the SLM-SRS system a stand-alone imaging platform.Results: We demonstrate that SLM-SRS microscopy permits rapid multiplexed SRS imaging of polystyrene and polymethyl methacrylate beads in Brownian motion in dimethyl sulfoxide (DMSO) at 70 ms intervals without motion artiacts. We further apply SLM-SRS to monitor the therapeautic effect of mild alkaline solution on cancer cells, which shows immediate apoptotic response. Finally, we visualize in vivo penetration of DMSO into the plant tissue and evaluate acute toxicity of DMSO on cellulose and proteins within the tissue.Conclusion: We develop novel SLM-SRS microscopy and affirm its broad applicability for rapid monitoring of dynamic biological processes at the subcellular and molecular level.