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Related Concept Videos

Total Internal Reflection Fluorescence Microscopy01:05

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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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Attenuated total reflectance (ATR) infrared spectroscopy is a powerful analytical technique used to study the composition of materials. It is widely employed in chemistry, materials science, forensic science, and other fields where sample characterization is required. ATR has several advantages over traditional transmission IR spectroscopy, including the requirement of little to no sample preparation and the ability to analyze a wide range of samples.
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Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
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Related Experiment Video

Updated: Aug 9, 2025

Total Internal Reflection Absorption Spectroscopy TIRAS for the Detection of Solvated Electrons at a Plasma-liquid Interface
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Surface Plasmon Resonance Microscopy Based on Total Internal Reflection.

Teliang Zhang1, Xueliang Wang1, Youjun Zeng2

  • 1Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen 518060, China.

Biosensors
|February 25, 2023
PubMed
Summary
This summary is machine-generated.

This study introduces a home-built total internal reflection surface plasmon resonance microscopy (TIR-SPRM) system. The enhanced system achieves 248 nm spatial resolution for imaging nanoparticles and monitoring biomolecular interactions.

Keywords:
biomolecular interactionsurface plasmon resonancesurface plasmon resonance microscopytotal internal reflection

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Area of Science:

  • Biophysics
  • Nanotechnology
  • Analytical Chemistry

Background:

  • Surface plasmon resonance microscopy (SPRM) is a powerful tool for label-free biological imaging.
  • High spatial resolution is crucial for analyzing nanoscale biological events.
  • Existing SPRM systems can be limited in resolution and nanoparticle imaging capabilities.

Purpose of the Study:

  • To develop and analyze a home-built total internal reflection SPRM (TIR-SPRM) system.
  • To improve the spatial resolution of SPRM for single nanoparticle imaging.
  • To demonstrate the system's capability in monitoring biomolecular interactions.

Main Methods:

  • Utilized a home-built TIR-SPRM system.
  • Analyzed the imaging principle of single nanoparticles.
  • Designed a ring filter and applied Fourier space deconvolution algorithm.
  • Measured specific binding between human IgG antigen and goat anti-human IgG antibody.

Main Results:

  • Achieved a spatial resolution of 248 nm by removing the parabolic tail of nanoparticle images.
  • Successfully imaged sparse nanoparticles.
  • Demonstrated the monitoring of specific biomolecular interactions, such as antigen-antibody binding.

Conclusions:

  • The developed TIR-SPRM system offers high spatial resolution for nanoscale imaging.
  • The system is capable of detecting and imaging sparse nanoparticles.
  • This SPRM approach effectively monitors biomolecular interactions in biological fields.