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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.

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Related Experiment Video

Updated: Jun 7, 2026

Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes
08:26

Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes

Published on: November 23, 2021

High-performance time-resolved fluorescence by direct waveform recording.

Joseph M Muretta1, Alexander Kyrychenko, Alexey S Ladokhin

  • 1Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, Minnesota 55455, USA.

The Review of Scientific Instruments
|November 2, 2010
PubMed
Summary
This summary is machine-generated.

A new high-performance time-resolved fluorescence (HPTRF) spectrometer enables rapid, precise fluorescence waveform acquisition. This advancement accelerates fluorescence spectroscopy, offering significant benefits for biological applications.

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Time-resolved Photophysical Characterization of Triplet-harvesting Organic Compounds at an Oxygen-free Environment Using an iCCD Camera
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Time-resolved Photophysical Characterization of Triplet-harvesting Organic Compounds at an Oxygen-free Environment Using an iCCD Camera

Published on: December 27, 2018

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Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes
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Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes

Published on: November 23, 2021

Time-resolved Photophysical Characterization of Triplet-harvesting Organic Compounds at an Oxygen-free Environment Using an iCCD Camera
06:08

Time-resolved Photophysical Characterization of Triplet-harvesting Organic Compounds at an Oxygen-free Environment Using an iCCD Camera

Published on: December 27, 2018

Area of Science:

  • Spectroscopy
  • Analytical Chemistry
  • Biophysics

Background:

  • Time-resolved fluorescence (TRF) spectroscopy is crucial for analyzing molecular dynamics and kinetics.
  • Conventional methods like time-correlated single photon counting (TCSPC) can be time-consuming, limiting throughput.
  • There is a need for faster, high-precision fluorescence measurement techniques.

Purpose of the Study:

  • To introduce a novel high-performance time-resolved fluorescence (HPTRF) spectrometer.
  • To demonstrate its capability for rapid and accurate acquisition of fluorescence emission waveforms.
  • To highlight its potential for high-throughput biological applications.

Main Methods:

  • Utilized an intense, high-repetition rate laser excitation source (1 µJ/pulse, 10 kHz, 1 ns FWHM).
  • Employed a transient digitizer with 0.125 ns per time point for waveform recording.
  • Acquired fluorescence decay curves for every laser pulse, enabling single-pulse measurements.

Main Results:

  • Achieved waveform acquisition rates at least 10^5 times faster than conventional TCSPC.
  • Obtained high signal-to-noise ratios (approx. 100) with single-pulse measurements.
  • Demonstrated high accuracy and precision in lifetime determination (down to 100 ps) and waveform linearity (within 1% error).
  • Successfully resolved components of samples with different lifetimes and mole fractions from data acquired in 0.1 s.

Conclusions:

  • The HPTRF spectrometer significantly enhances the speed and efficiency of time-resolved fluorescence measurements.
  • It provides high accuracy and precision comparable to established methods.
  • The instrument opens new possibilities for high-throughput biological studies, including transient kinetics and multidimensional fluorescence analysis.