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Related Concept Videos

Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

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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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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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Two-dimensional (2D) microscopy encompasses a range of optical techniques that capture images within a single focal plane, offering detailed representations of microscopic structures. These techniques are essential in biological and medical research, enabling the visualization of cellular and subcellular structures with different levels of contrast and specificity.There are several major types of 2D microscopy, each with strengths and applications.Bright-Field MicroscopyBright-field microscopy...
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Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
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Photoluminescence offers a wide range of applications due to its inherent sensitivity and selectivity. This technique allows for both direct and indirect analyses of the analyte. Direct quantitative analysis is possible when the analyte exhibits a favorable quantum yield for fluorescence or phosphorescence. However, an indirect analysis may be feasible if the analyte is not fluorescent or phosphorescent, or if the quantum yield is unfavorable. Indirect methods include reacting the analyte with...
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Related Experiment Video

Updated: Apr 25, 2026

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
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High-speed fluorescence lifetime imaging microscopy: techniques, applications, and prospects.

Hiroshi Kanno1,2, Fan Li1, Jongchan Park3

  • 1University of Tokyo, School of Science, Department of Chemistry, Tokyo, Japan.

Biophotonics Discovery
|April 24, 2026
PubMed
Summary

High-speed Fluorescence Lifetime Imaging Microscopy (FLIM) overcomes previous speed limitations. This advancement enables faster cellular dynamics observation and broadens biomedical applications.

Keywords:
fluorescence lifetimefluorescence lifetime imaging microscopyhigh-speed imaging

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Area of Science:

  • Biomedical Imaging
  • Microscopy Techniques
  • Cellular Dynamics

Background:

  • Fluorescence Lifetime Imaging Microscopy (FLIM) offers quantitative measurements superior to intensity-based methods.
  • Slow imaging speed has historically limited FLIM's biological and medical applications.

Purpose of the Study:

  • To review recent advancements in high-speed FLIM techniques.
  • To explore the expanded biomedical applications and future potential of accelerated FLIM.

Main Methods:

  • Categorization of high-speed FLIM into time-domain and frequency-domain approaches.
  • Discussion of wide-field and beam-scanning imaging schemes, including their combinations.

Main Results:

  • Innovations in fluorescence detection and excitation strategies have significantly boosted FLIM speed.
  • High-speed FLIM now allows observation of rapid cellular processes like neural spiking.
  • Feasible large-scale analysis of heterogeneous cells and potential for video-rate 3D FLIM.

Conclusions:

  • High-speed FLIM development opens new avenues in cell biology, biophysics, and neurology.
  • Future directions include fit-free analysis, 3D FLIM, and FLIM-based cell sorting.