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Related Concept Videos

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.
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,...
Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

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Related Experiment Video

Updated: Jun 24, 2026

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
09:45

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Published on: February 9, 2012

Stroboscopic fluorescence lifetime imaging.

Mark D Holton1, Oscar R Silvestre, Rachel J Errington

  • 1School of Medicine, Swansea University, Singleton Park, Swansea, UK.

Optics Express
|April 1, 2009
PubMed
Summary
This summary is machine-generated.

We developed a new fluorescence lifetime imaging method using periodic optical excitation, enabling real-time cell imaging without high-speed detectors. This technique tracks quantum dot endocytosis by cancer cells, revealing cellular environment changes.

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Last Updated: Jun 24, 2026

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
09:45

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Published on: February 9, 2012

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Published on: April 7, 2023

Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy
09:30

Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy

Published on: January 18, 2017

Area of Science:

  • Biophotonics
  • Cellular Imaging
  • Quantum Dot Research

Background:

  • Fluorescence lifetime imaging (FLIM) traditionally requires high temporal resolution for detection.
  • Probing cellular dynamics often necessitates advanced imaging techniques.
  • Quantum dots (QDs) are valuable probes in cellular research but their optical properties can change in vivo.

Purpose of the Study:

  • To introduce a novel fluorescence lifetime imaging technique that bypasses the need for time-resolved detectors.
  • To demonstrate the capability of this technique for real-time, wide-field imaging of cellular processes.
  • To validate the method by observing the endocytosis of quantum dots in a cancer cell line.

Main Methods:

  • Utilizing a time-integrated response to periodic optical excitation (stroboscopic method).
  • Employing a Dirac pulse train to probe the frequency response, linked to fluorophore relaxation rates.
  • Implementing the technique with standard CCD cameras for wide-field, real-time acquisition.

Main Results:

  • Successfully imaged the endocytosis of inorganic quantum dots in a cancer cell line in real-time.
  • Observed a significant change in quantum dot fluorescence lifetime from ~40 ns to <10 ns due to intracellular surface charging.
  • Achieved a temporal resolution of 15 ns, half the excitation period.

Conclusions:

  • The developed stroboscopic FLIM technique enables video-rate lifetime imaging using standard equipment.
  • This method is suitable for studying millisecond-scale cellular dynamics and high-throughput screening.
  • The observed lifetime shift in QDs provides insights into the intracellular environment.