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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.
Attenuated Total Reflectance (ATR) Infrared Spectroscopy: Overview01:13

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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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Phase Contrast and Differential Interference Contrast Microscopy01:26

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Phase-Contrast Microscopes
In-phase-contrast microscopes, interference between light directly passing through a cell and light refracted by cellular components is used to create high-contrast, high-resolution images without staining. It is the oldest and simplest type of microscope that creates an image by altering the wavelengths of light rays passing through the specimen. Altered wavelength paths are created using an annular stop in the condenser. The annular stop produces a hollow cone of...

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Related Experiment Video

Updated: Jul 4, 2026

Digital Inline Holographic Microscopy (DIHM) of Weakly-scattering Subjects
10:16

Digital Inline Holographic Microscopy (DIHM) of Weakly-scattering Subjects

Published on: February 8, 2014

Digital holography of total internal reflection.

William M Ash1, Myung K Kim

  • 1Digital Holography and Microscopy Laboratory, Dept. of Physics, University of South Florida, Tampa, FL 33620, USA. wash@mail.usf.edu

Optics Express
|June 26, 2008
PubMed
Summary

We developed total internal reflection holographic microscopy (TIRHM) for surface imaging. This new technique uses quantitative phase microscopy to reveal material properties near reflective surfaces.

Area of Science:

  • Optics and Photonics
  • Microscopy
  • Materials Science

Background:

  • Quantitative phase microscopy (QPM) provides label-free imaging of transparent specimens.
  • Digital holography is a powerful QPM technique capable of reconstructing both amplitude and phase information.
  • Total internal reflection (TIR) is sensitive to the evanescent field near an interface, enabling surface-specific measurements.

Purpose of the Study:

  • To introduce and characterize a novel microscopy technique, total internal reflection holographic microscopy (TIRHM).
  • To leverage digital holography for quantitative phase imaging under total internal reflection conditions.
  • To demonstrate the capability of TIRHM for analyzing materials at or near reflective surfaces.

Main Methods:

  • Development of the total internal reflection holographic microscopy (TIRHM) system.

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  • Application of quantitative phase microscopy (QPM) using digital holography to capture phase profiles.
  • Theoretical modeling of the imaging characteristics of TIRHM.
  • Experimental validation of the imaging capabilities of TIRHM.

    • Main Results:

    • Successful implementation of TIRHM, combining TIR with holographic QPM.
    • Demonstration of phase profile imaging modulated by surface-bound materials.
    • Theoretical models accurately predict TIRHM imaging behavior.
    • Experimental results confirm the practical imaging capabilities of TIRHM for surface analysis.

    Conclusions:

    • TIRHM is a viable new microscopy technique for surface and near-surface material characterization.
    • The technique offers quantitative phase information in a surface-sensitive manner.
    • TIRHM holds promise for applications requiring high-resolution imaging of interfacial phenomena.