Toward photoswitchable electronic pre-resonance stimulated Raman probes

  • Dongkwan Lee
    Division of Chemistry and Chemical Engineering, California Institute of Technology 1 , Pasadena, California 91125, USA
  • Chenxi Qian
    Division of Chemistry and Chemical Engineering, California Institute of Technology 1 , Pasadena, California 91125, USA
  • Haomin Wang
    Division of Chemistry and Chemical Engineering, California Institute of Technology 1 , Pasadena, California 91125, USA
  • Lei Li
    Division of Engineering and Applied Science, California Institute of Technology 2 , Pasadena, California 91125, USA
  • Kun Miao
    Division of Chemistry and Chemical Engineering, California Institute of Technology 1 , Pasadena, California 91125, USA
  • Jiajun Du
    Division of Chemistry and Chemical Engineering, California Institute of Technology 1 , Pasadena, California 91125, USA
  • Daria M. Shcherbakova
    Department of Anatomy and Structural Biology, and Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine 3 , Bronx, New York 10461, USA
  • Vladislav V. Verkhusha
    Department of Anatomy and Structural Biology, and Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine 3 , Bronx, New York 10461, USA
  • Lihong V. Wang
    Division of Engineering and Applied Science, California Institute of Technology 2 , Pasadena, California 91125, USA
  • Lu Wei
    Division of Chemistry and Chemical Engineering, California Institute of Technology 1 , Pasadena, California 91125, USA

抄録

<jats:p>Reversibly photoswitchable probes allow for a wide variety of optical imaging applications. In particular, photoswitchable fluorescent probes have significantly facilitated the development of super-resolution microscopy. Recently, stimulated Raman scattering (SRS) imaging, a sensitive and chemical-specific optical microscopy, has proven to be a powerful live-cell imaging strategy. Driven by the advances of newly developed Raman probes, in particular the pre-resonance enhanced narrow-band vibrational probes, electronic pre-resonance SRS (epr-SRS) has achieved super-multiplex imaging with sensitivity down to 250 nM and multiplexity up to 24 colors. However, despite the high demand, photoswitchable Raman probes have yet to be developed. Here, we propose a general strategy for devising photoswitchable epr-SRS probes. Toward this goal, we exploit the molecular electronic and vibrational coupling, in which we switch the electronic states of the molecules to four different states to turn their ground-state epr-SRS signals on and off. First, we showed that inducing transitions to both the electronic excited state and triplet state can effectively diminish the SRS peaks. Second, we revealed that the epr-SRS signals can be effectively switched off in red-absorbing organic molecules through light-facilitated transitions to a reduced state. Third, we identified that photoswitchable proteins with near-infrared photoswitchable absorbance, whose states are modulable with their electronic resonances detunable toward and away from the pump photon energy, can function as the photoswitchable epr-SRS probes with desirable sensitivity (&lt;1 µM) and low photofatigue (&gt;40 cycles). These photophysical characterizations and proof-of-concept demonstrations should advance the development of novel photoswitchable Raman probes and open up the unexplored Raman imaging capabilities.</jats:p>

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