Higher-order coherent anti-Stokes Raman scattering microscopy realizes label-free super-resolution vibrational imaging

被引:105
作者
Gong, Li [1 ]
Zheng, Wei [1 ]
Ma, Ying [1 ]
Huang, Zhiwei [1 ]
机构
[1] Natl Univ Singapore, Fac Engn, Dept Biomed Engn, Opt Bioimaging Lab, Singapore, Singapore
基金
英国医学研究理事会;
关键词
RESOLUTION LIMIT; IN-VIVO; PHOTODAMAGE; SPECTRA;
D O I
10.1038/s41566-019-0535-y
中图分类号
O43 [光学];
学科分类号
070207 ; 0803 ;
摘要
Higher-order (fifth and seventh order) coherent anti-Stokes Raman scattering microscopy is demonstrated to break the diffraction limit for label-free super-resolution vibrational imaging for live cells such as HeLa and buccal cells. Coherent Raman scattering (for example, coherent anti-Stokes Raman scattering (CARS) and stimulated Raman scattering) microscopy has emerged as a powerful tool for label-free biomolecular imaging in biological and biomedical systems, but its spatial resolution is diffraction limited. Here, we report a higher-order coherent anti-Stokes Raman scattering (HO-CARS) microscopy to break the diffraction limit for label-free, super-resolution vibrational imaging. The resolution enhancement of HO-CARS microscopy has been analysed and demonstrated in biological samples (for example, live HeLa and buccal cells). The HO-CARS technique provides an inherent high resonant to non-resonant background ratio compared with conventional CARS microscopy. We affirm that under a tight focusing, the HO-CARS signal originating from the higher-order nonlinear process (chi((5)), chi((7))) dominates over the cascaded lower-order nonlinear process (chi((3))), yielding much richer spectroscopic information. This study illustrates that HO-CARS microscopy can be an appealing tool for label-free, super-resolution imaging in biological and biomedical systems with high image contrast.
引用
收藏
页码:115 / +
页数:9
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