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Related Concept Videos

Total Internal Reflection Fluorescence Microscopy01:05

Total Internal Reflection Fluorescence Microscopy

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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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Super-resolution Fluorescence Microscopy01:37

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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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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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Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

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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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Internal Energy02:00

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The total of all possible kinds of energy present in a substance is called the internal energy (U), sometimes symbolized as E. Suppose a system with initial internal energy, Uinitial, undergoes a change in energy (transfer of work or heat), and the final internal energy of the system is Ufinal. Change in internal energy equals the difference between Ufinal and Uinitial.
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Internal Receptors01:31

Internal Receptors

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Many cellular signals are hydrophilic and therefore cannot pass through the plasma membrane. However, small or hydrophobic signaling molecules can cross the hydrophobic core of the plasma membrane and bind to internal, or intracellular, receptors that reside within the cell. Many mammalian steroid hormones use this mechanism of cell signaling, as does nitric oxide (NO) gas.
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Related Experiment Video

Updated: Feb 10, 2026

Visualizing Adhesion Formation in Cells by Means of Advanced Spinning Disk-Total Internal Reflection Fluorescence Microscopy
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Visualizing Adhesion Formation in Cells by Means of Advanced Spinning Disk-Total Internal Reflection Fluorescence Microscopy

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Single-shot super-resolution total internal reflection fluorescence microscopy.

Min Guo1, Panagiotis Chandris2, John Paul Giannini2,3

  • 1Section on High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD, USA. min.guo@nih.gov.

Nature Methods
|May 9, 2018
PubMed
Summary
This summary is machine-generated.

Researchers developed instant TIRF-SIM, a super-resolution microscopy technique. This method enhances imaging resolution near surfaces without sacrificing speed, enabling faster, clearer live-cell studies.

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Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins
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Area of Science:

  • Biophysics
  • Microscopy
  • Cell Biology

Background:

  • Total internal reflection fluorescence microscopy (TIRFM) is crucial for live-cell imaging near surfaces.
  • Standard TIRFM resolution limits detailed observation of subcellular structures.
  • Faster imaging techniques are needed to capture dynamic cellular processes.

Purpose of the Study:

  • To enhance the lateral spatial resolution of TIRFM.
  • To develop a super-resolution microscopy technique compatible with live samples.
  • To achieve high-speed, high-contrast imaging near the coverslip surface.

Main Methods:

  • Combined instant structured illumination microscopy (iSIM) with TIRFM to create instant TIRF-SIM.
  • Applied the technique to live biological samples.
  • Optimized imaging parameters for speed and resolution.

Main Results:

  • Achieved a lateral spatial resolution of 115 ± 13 nm.
  • Maintained high imaging speed, with frame rates up to 100 Hz.
  • Enabled rapid, high-contrast super-resolution imaging of live samples near the coverslip.

Conclusions:

  • Instant TIRF-SIM significantly improves TIRFM's spatial resolution.
  • The technique allows for fast, super-resolution live-cell imaging.
  • This advancement opens new possibilities for studying dynamic cellular events at high resolution.