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Related Concept Videos

Total Internal Reflection Fluorescence Microscopy01:05

Total Internal Reflection Fluorescence Microscopy

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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Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...
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Related Experiment Video

Updated: Jun 7, 2026

High-Throughput Total Internal Reflection Fluorescence and Direct Stochastic Optical Reconstruction Microscopy Using a Photonic Chip
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Superresolution confocal technology for displacement measurements based on total internal reflection.

Cuifang Kuang1, M Yakut Ali, Xiang Hao

  • 1State Key Laboratory of Modern Optical Instrumentations, Zhejiang University, Hangzhou 310027, China. cfkuang@zju.edu.cn

The Review of Scientific Instruments
|November 2, 2010
PubMed
Summary

A new method using total internal reflection filters and confocal microscopy achieves sub-nanometer axial resolution for displacement measurements. This technique significantly enhances measurement accuracy and stability compared to traditional methods.

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Area of Science:

  • Optics
  • Metrology
  • Microscopy

Background:

  • Accurate displacement measurement is crucial in various scientific and industrial fields.
  • Existing methods often face limitations in achieving high axial resolution.
  • Enhancing the resolution of nonlinearity in reflectance curves is key for precise measurements.

Purpose of the Study:

  • To propose and validate a novel method for high-resolution axial displacement measurement.
  • To theoretically analyze the principles of the proposed measurement technique.
  • To experimentally verify the feasibility, accuracy, and stability of the developed system.

Main Methods:

  • Development of a novel measurement method combining total internal reflection filters and confocal microscopy principles.
  • Theoretical analysis of the fundamental measurement principles and the confocal detection scheme.
  • Construction of a prototype system and performance evaluation through laboratory experiments.

Main Results:

  • The proposed confocal detection scheme effectively enhances the resolution of the reflectance curve's nonlinearity.
  • Experimental results demonstrate an axial resolution better than 1 nm within a 200 nm range.
  • The developed system shows superior performance, achieving threefold improvement over plane reflector methods.

Conclusions:

  • The novel method based on total internal reflection filters and confocal microscopy offers superior axial resolution for displacement measurements.
  • The developed prototype system is feasible, accurate, and stable for high-precision metrology.
  • This technique represents a significant advancement in achieving sub-nanometer displacement resolution.