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Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
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Advances in NIR-II Fluorescent Nanoprobes: Design Principles, Optical Engineering, and Emerging Translational

Nargish Parvin1, Mohammad Aslam2, Md Najib Alam1

  • 1School of Mechanical Engineering, Yeungnam University, Gyeongsan 38541, Republic of Korea.

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Near-infrared-II (NIR-II) fluorescent nanoprobes offer deep-tissue imaging with high resolution. This review details advances in quantum dots, carbon dots, upconversion nanoparticles, and silica nanoparticles for biomedical applications.

Keywords:
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Area of Science:

  • Biomedical Engineering
  • Materials Science
  • Optical Imaging

Background:

  • Fluorescent nanoprobes operating in the NIR-II window are crucial for advanced biomedical imaging.
  • Their properties include deep-tissue penetration, reduced scattering, and high spatial resolution.
  • These nanoprobes facilitate sensitive detection and targeted visualization of biological structures in vivo.

Purpose of the Study:

  • To review recent advances in the design and optical engineering of NIR-II nanoprobes.
  • To highlight four key nanoprobe families: quantum dots, carbon dots, upconversion nanoparticles, and dye-doped silica nanoparticles.
  • To discuss emerging strategies for probe construction and their translational potential.

Main Methods:

  • Review of recent literature on NIR-II nanoprobes.
  • Analysis of design strategies and optical engineering advancements.
  • Focus on activatable, targeted, and ratiometric probe construction.

Main Results:

  • Detailed examination of four major NIR-II nanoprobe families.
  • Discussion of strategies for enhancing imaging performance and multifunctionality.
  • Summary of progress in biosafety, synthesis, stability, and preclinical validation.

Conclusions:

  • Engineered NIR-II nanoprobes show significant progress toward practical use in biomedical imaging.
  • Further research is needed to address challenges influencing clinical readiness.
  • These nanoprobes are advancing precision diagnostics and in vivo visualization.