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An Integrated System to Remotely Trigger Intracellular Signal Transduction by Upconversion Nanoparticle-mediated Kinase Photoactivation
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Multi-photon near-infrared emission saturation nanoscopy using upconversion nanoparticles.

Chaohao Chen1, Fan Wang2, Shihui Wen1

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|August 19, 2018
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Upconversion nanoparticles enable near-infrared emission saturation nanoscopy for deep tissue super-resolution imaging. This novel method achieves sub-50nm resolution in thick tissues with significantly lower excitation intensity than conventional techniques.

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Area of Science:

  • Biophotonics
  • Nanotechnology
  • Super-resolution microscopy

Background:

  • Multiphoton fluorescence microscopy (MPF) offers deep tissue penetration and reduced phototoxicity using near-infrared excitation.
  • Stimulated emission depletion (STED) microscopy can overcome diffraction limits but requires complex laser setups and inefficient probes.
  • Existing methods face limitations in achieving high-resolution imaging deep within biological tissues.

Purpose of the Study:

  • To develop a simplified super-resolution microscopy technique for deep tissue imaging.
  • To utilize upconversion nanoparticles (UCNPs) for enhanced near-infrared emission.
  • To overcome the limitations of conventional multiphoton and STED microscopy for in vivo applications.

Main Methods:

  • Employing upconversion nanoparticles (UCNPs) as novel fluorescent probes.
  • Utilizing near-infrared emission saturation (NIRES) nanoscopy.
  • Implementing a doughnut beam excitation with a 980 nm diode laser and detecting emission at 800 nm.
  • Imaging through 93 μm thick liver tissue.

Main Results:

  • Achieved sub-50 nm resolution, approximately 1/20th of the excitation wavelength.
  • Demonstrated super-resolution imaging of single UCNPs within thick biological tissue.
  • Required excitation intensity orders of magnitude lower than conventional multiphoton microscopy dyes.
  • Successfully imaged through 93 μm of liver tissue.

Conclusions:

  • Upconversion nanoparticles unlock a new mode of nanoscopy (NIRES) for deep tissue super-resolution imaging.
  • This method provides a simplified approach for high-resolution imaging in scattering biological tissues.
  • The technique shows promise for advanced applications like single molecule tracking in deep tissues.