Department of Chemistry, Columbia University, New York, NY, 10027, USA
Wei Min (wm2256@columbia.edu)
纸质出版日期:2021-05-31,
网络出版日期:2021-04-20,
收稿日期:2020-12-29,
修回日期:2021-03-24,
录用日期:2021-03-26
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Super-resolution vibrational microscopy by stimulated Raman excited fluorescence[J]. LSA, 2021,10(5):906-915.
Xiong, H. Q. et al. Super-resolution vibrational microscopy by stimulated Raman excited fluorescence. Light: Science & Applications, 10, 906-915 (2021).
Super-resolution vibrational microscopy by stimulated Raman excited fluorescence[J]. LSA, 2021,10(5):906-915. DOI: 10.1038/s41377-021-00518-5.
Xiong, H. Q. et al. Super-resolution vibrational microscopy by stimulated Raman excited fluorescence. Light: Science & Applications, 10, 906-915 (2021). DOI: 10.1038/s41377-021-00518-5.
Inspired by the revolutionary impact of super-resolution fluorescence microscopy
super-resolution Raman imaging has been long pursued because of its much higher chemical specificity than the fluorescence counterpart. However
vibrational contrasts are intrinsically less sensitive compared with fluorescence
resulting in only mild resolution enhancement beyond the diffraction limit even with strong laser excitation power. As such
it is still a great challenge to achieve biocompatible super-resolution vibrational imaging in the optical far-field. In 2019 Stimulated Raman Excited Fluorescence (SREF) was discovered as an ultrasensitive vibrational spectroscopy that combines the high chemical specificity of Raman scattering and the superb sensitivity of fluorescence detection. Herein we developed a novel super-resolution vibrational imaging method by harnessing SREF as the contrast mechanism. We first identified the undesired role of anti-Stokes fluorescence background in preventing direct adoption of super-resolution fluorescence technique. We then devised a frequency-modulation (FM) strategy to remove the broadband backgrounds and achieved high-contrast SREF imaging. Assisted by newly synthesized SREF dyes
we realized multicolor FM-SREF imaging with nanometer spectral resolution. Finally
by integrating stimulated emission depletion (STED) with background-free FM-SREF
we accomplished high-contrast super-resolution vibrational imaging with STED-FM-SREF whose spatial resolution is only determined by the signal-to-noise ratio. In our proof-of-principle demonstration
more than two times of resolution improvement is achieved in biological systems with moderate laser excitation power
which shall be further refined with optimized instrumentation and imaging probes. With its super resolution
high sensitivity
vibrational contrast
and mild laser excitation power
STED-FM-SREF microscopy is envisioned to aid a wide variety of applications.
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