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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.
Overview of Microscopy Techniques01:22

Overview of Microscopy Techniques

The early pioneers of microscopy opened a window into the invisible world of microorganisms. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes that leveraged nonvisible light, such as fluorescence microscopy that uses an ultraviolet light source and electron microscopy that uses short-wavelength electron beams. These advances significantly improved magnification, image resolution, and contrast. By comparison, the...
Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

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,...
Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

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 developed.
Three-Dimensional Microscopy in Microbiology01:28

Three-Dimensional Microscopy in Microbiology

Three-dimensional imaging techniques are essential in cell biology, allowing researchers to visualize intricate cellular structures with high resolution. Two prominent methods, Differential Interference Contrast Microscopy (DIC) and Confocal Scanning Laser Microscopy (CSLM), provide distinct advantages for imaging live and thick specimens, respectively.Differential Interference Contrast MicroscopyDIC microscopy enhances contrast in transparent, unstained samples by converting phase...
Electron Microscope Tomography and Single-particle Reconstruction01:07

Electron Microscope Tomography and Single-particle Reconstruction

Transmission electron microscopy (TEM) can be used to determine the 3D structure of biological samples with the help of techniques such as electron microscope tomography and single-particle reconstruction. While single-particle reconstruction can examine macromolecules and macromolecular complexes in vitro conditions only, tomography permits the study of cell components or small cells in vivo.
Electron Tomography
Electron tomography can be performed either in TEM or STEM (scanning transmission...

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Related Experiment Video

Updated: Jul 3, 2026

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins
06:43

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins

Published on: May 3, 2022

Combined scanning probe and total internal reflection fluorescence microscopy.

John Oreopoulos1, Christopher M Yip

  • 1Institute of Biomaterials and Biomedical Engineering, Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, 160 College St, Toronto, Ont., Canada M5S 3E1.

Methods (San Diego, Calif.)
|July 8, 2008
PubMed
Summary
This summary is machine-generated.

This study combines scanning probe and optical microscopy for real-time imaging of biomolecules. This powerful platform reveals molecular structure and dynamics, especially interactions at membrane surfaces.

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Last Updated: Jul 3, 2026

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins
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High-Throughput Total Internal Reflection Fluorescence and Direct Stochastic Optical Reconstruction Microscopy Using a Photonic Chip

Published on: November 16, 2019

Area of Science:

  • Biophysics
  • Microscopy
  • Biochemistry

Background:

  • Investigating biomolecular structure-function relationships and dynamics requires advanced imaging techniques.
  • Simultaneous multi-modal imaging offers deeper insights into molecular behavior in situ and in real-time.

Purpose of the Study:

  • To describe strategies for coupling scanning probe microscopy (SPM) with total internal reflection fluorescence microscopy (TIRFM).
  • To highlight the potential applications of this integrated platform for studying biomolecular interactions, particularly at membrane surfaces.

Main Methods:

  • Integration of scanning probe microscopy (SPM) with total internal reflection fluorescence microscopy (TIRFM).
  • Utilizing complementary techniques like optical microscopy and spectroscopy for simultaneous characterization.
  • Achieving three-dimensional imaging of individual molecules with nanometer resolution.

Main Results:

  • Demonstration of a combined SPM-TIRFM platform for high-resolution biomolecular imaging.
  • Characterization of molecular structure, interactions, and dynamics at membrane interfaces.
  • Identification of practical strategies and challenges in implementing such coupled microscopy systems.

Conclusions:

  • The coupled SPM-TIRFM platform provides a powerful tool for real-time, in situ investigation of biomolecular interactions.
  • This technology enables nanometer-resolution imaging and simultaneous functional characterization.
  • Significant potential for advancing our understanding of biological processes at membrane surfaces.