Microsecond scale vibrational spectroscopic imaging by multiplex stimulated Raman scattering microscopy
Microsecond scale vibrational spectroscopic imaging by multiplex stimulated Raman scattering microscopy
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DOI：
10.1038/lsa.2015.38
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发表时间：
2015-03-01
影响因子：
19.4
通讯作者：
Cheng, Ji-Xin
中科院分区：
文献类型：
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作者：
Liao, Chien-Sheng;Slipchenko, Mikhail N.;Cheng, Ji-Xin
实时振动光谱成像是用于监测细胞状态和细胞过程的无标签方式。高动态系统的拉曼光谱成像受到毫秒到秒尺度上相对缓慢的光谱采集的抑制。在这里，我们报告了微秒尺度的振动光谱成像，通过锁定自由平行检测光谱分散的受激拉曼散射信号。使用自制的调谐放大器阵列，我们的方法能够在宽带脉冲定义的窗口内以32 μ s的速度进行拉曼光谱采集，并且具有接近射击噪声限制的检测灵敏度。结合多元曲线分辨率分析，我们的平台可以实现单个活细胞中脂滴的成分定位、细胞内类视黄醇代谢的观察、秀丽隐杆线虫中富含蛋白质的细胞器中脂滴的区分、快速流动肿瘤细胞的光谱检测以及体内皮肤组织中药物扩散的监测。所报道的技术为显微镜下细胞室的成分分析和流式细胞仪下单细胞的高通量光谱分析开辟了新的机会。
Real-time vibrational spectroscopic imaging is desired for monitoring cellular states and cellular processes in a label-free manner. Raman spectroscopic imaging of highly dynamic systems is inhibited by relatively slow spectral acquisition on millisecond to second scale. Here, we report microsecond scale vibrational spectroscopic imaging by lock-in free parallel detection of spectrally dispersed stimulated Raman scattering signal. Using a homebuilt tuned amplifier array, our method enables Raman spectral acquisition, within the window defined by the broadband pulse, at the speed of 32 mu s and with close to shot-noise limited detection sensitivity. Incorporated with multivariate curve resolution analysis, our platform allows compositional mapping of lipid droplets in single live cells, observation of intracellular retinoid metabolism, discrimination of fat droplets from protein-rich organelles in Caenorhabditis elegans, spectral detection of fast flowing tumor cells and monitoring drug diffusion through skin tissue in vivo. The reported technique opens new opportunities for compositional analysis of cellular compartment in a microscope setting and high-throughput spectral profiling of single cells in a flow cytometer setting.