High-resolution optical microscopes that can break 180 nm in spatial resolution set to conventional microscopies are much-needed tools.However,current optical microscopes have to rely on exogenous fluorescent labels t...High-resolution optical microscopes that can break 180 nm in spatial resolution set to conventional microscopies are much-needed tools.However,current optical microscopes have to rely on exogenous fluorescent labels to achieve high resolution in biological imaging.Herein,we report near-resonance enhanced label-free stimulated Raman scattering(SRS)microscopy with a lateral resolution near 130 nm,in which the high-resolution image contrast originates directly from a low concentration of endogenous biomolecules,with sensitivity gains of approximately 23 times.Moreover,by using a 0.3-m-long optical fiber,we developed hyperspectral SRS microscopy based on spectral focusing technology.Attributed to enhancements in spatial resolution and sensitivity,we demonstrated highresolution imaging of three-dimensional structures in single cells and high-resolution mapping of large-scale intact mouse brain tissues in situ.By using enhanced high-resolution hyperspectral SRS,we chemically observed sphingomyelin distributed in the myelin sheath that insulates single axons.Our concept opens the door to biomedical imaging with~130 nm resolution.展开更多
We designed a lysosome-selective Raman probe by conjugating bisphenylbutadiyne with morpholine, a well-known lysosome targeting moiety. This probe, named Lyso-BADY, has a Raman peak 28 times more intense than that of ...We designed a lysosome-selective Raman probe by conjugating bisphenylbutadiyne with morpholine, a well-known lysosome targeting moiety. This probe, named Lyso-BADY, has a Raman peak 28 times more intense than that of 5-ethynyl-2'-deoxyuridine. Lysosome in living cells was successfully visualized by hyperspectral stimulated Raman scattering (SRS) microscopy with this extracellular probe. Further study showed that the Raman signal of Lyso-BADY remained steady and strong even after a prolonged irradiation time. The photo-stability feature of Lyso-BADY rendered molecules of the similar structure as potentially versatile probe for continuous imaging in the future.展开更多
基金support from the National Key Research and Development Program of China(2016YFA0201403)National Natural Science Foundation of China(61675075)+3 种基金Science Fund for Creative Research Group of China(61421064)Chinese Recruitment Program of Global ExpertsDirector Fund of the Wuhan National Laboratory for Optoelectronicsby the Natural Science Foundation of Hubei Province(grant 2017CFB591).
文摘High-resolution optical microscopes that can break 180 nm in spatial resolution set to conventional microscopies are much-needed tools.However,current optical microscopes have to rely on exogenous fluorescent labels to achieve high resolution in biological imaging.Herein,we report near-resonance enhanced label-free stimulated Raman scattering(SRS)microscopy with a lateral resolution near 130 nm,in which the high-resolution image contrast originates directly from a low concentration of endogenous biomolecules,with sensitivity gains of approximately 23 times.Moreover,by using a 0.3-m-long optical fiber,we developed hyperspectral SRS microscopy based on spectral focusing technology.Attributed to enhancements in spatial resolution and sensitivity,we demonstrated highresolution imaging of three-dimensional structures in single cells and high-resolution mapping of large-scale intact mouse brain tissues in situ.By using enhanced high-resolution hyperspectral SRS,we chemically observed sphingomyelin distributed in the myelin sheath that insulates single axons.Our concept opens the door to biomedical imaging with~130 nm resolution.
基金the National Natural Science Foundation of China (Nos. 21432008, 91753201 and 21721005) for financial support
文摘We designed a lysosome-selective Raman probe by conjugating bisphenylbutadiyne with morpholine, a well-known lysosome targeting moiety. This probe, named Lyso-BADY, has a Raman peak 28 times more intense than that of 5-ethynyl-2'-deoxyuridine. Lysosome in living cells was successfully visualized by hyperspectral stimulated Raman scattering (SRS) microscopy with this extracellular probe. Further study showed that the Raman signal of Lyso-BADY remained steady and strong even after a prolonged irradiation time. The photo-stability feature of Lyso-BADY rendered molecules of the similar structure as potentially versatile probe for continuous imaging in the future.
