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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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Fluorescence Lifetime Macro Imager for Biomedical Applications
06:01

Fluorescence Lifetime Macro Imager for Biomedical Applications

Published on: April 7, 2023

Frequency Multiplexed In Vivo Multiphoton Phosphorescence Lifetime Microscopy.

Scott S Howard1, Adam Straub, Nicholas Horton

  • 1School of Applied and Engineering Physics, Cornell University, 212 Clark Hall, Ithaca, NY 14853. Department of Electrical Engineering, University of Notre Dame, 275 Fitzpatrick Hall, Notre Dame, IN 46556.

Nature Photonics
|March 9, 2013
PubMed
Summary

This study introduces a faster multiphoton microscopy-phosphorescence lifetime imaging microscopy (MPM-PLIM) system. The enhanced system achieves 100x higher pixel rates for in vivo imaging of oxygen levels in scattering tissue.

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Published on: September 30, 2021

Area of Science:

  • Biomedical Optics
  • Microscopy Techniques
  • In Vivo Imaging

Background:

  • Multiphoton microscopy (MPM) enables deep optical sectioning in scattering tissues.
  • Phosphorescence lifetime imaging microscopy (PLIM) provides chemical information via Förster resonance energy transfer and phosphorescence quenching.
  • Current point-measurement PLIM limits imaging speed due to fundamental constraints on fluorescence generation rate.

Purpose of the Study:

  • To overcome the speed limitations of conventional PLIM systems.
  • To develop a parallelized MPM-PLIM system for enhanced imaging speed and depth.
  • To enable high-resolution, real-time in vivo imaging of physiological parameters.

Main Methods:

  • Development and implementation of a parallel-excitation/parallel-collection MPM-PLIM system.
  • Simultaneous acquisition of lifetime and intensity images at depth in vivo.
  • Demonstration of full-frame three-dimensional in vivo PLIM imaging using phosphorescent quenching dye.

Main Results:

  • Achieved a 100-fold increase in pixel rate compared to conventional MPM-PLIM configurations.
  • Successfully acquired simultaneous lifetime and intensity images deep within scattering tissue in vivo.
  • Presented the first full-frame 3D in vivo PLIM imaging of phosphorescent quenching dye.

Conclusions:

  • The parallelized MPM-PLIM system significantly enhances imaging speed for in vivo applications.
  • This technology enables new possibilities for studying oxygen partial pressure and other chemical dynamics in biological tissues.
  • The developed system establishes a novel platform for advanced biological and medical imaging.