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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 4, 2026

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy
12:51

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Published on: December 9, 2013

Simultaneous multiple-excitation multiphoton microscopy yields increased imaging sensitivity and specificity.

Margaret T Butko1, Mikhail Drobizhev, Nikolay S Makarov

  • 1Department of Neuroscience and Biomedical Graduate Program, Howard Hughes Medical Institute, University of California, San Diego, La Jolla, CA 92093, USA.

BMC Biotechnology
|March 4, 2011
PubMed
Summary
This summary is machine-generated.

Multiphoton microscopy (MPM) challenges in multi-color imaging are overcome with a novel multiple-excitation MPM (ME-MPM) system. This system uses two independent light sources to selectively excite fluorophores, improving signal quality and reducing bleed-through for biological samples.

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Published on: October 28, 2018

Area of Science:

  • Biophotonics
  • Microscopy
  • Molecular Imaging

Background:

  • Multiphoton microscopy (MPM) offers advantages like 3D resolution and deeper tissue imaging compared to conventional microscopy.
  • Multi-color imaging in MPM is challenging due to overlapping two-photon absorption peaks of fluorophores.
  • Current methods using single excitation wavelengths lead to inefficient and unequal dye excitation.

Purpose of the Study:

  • To develop a novel multiphoton microscopy system for improved multi-color imaging.
  • To address the limitations of single-excitation multiphoton microscopy in differentiating fluorophores.

Main Methods:

  • Developed a multiple-excitation multiphoton microscopy (ME-MPM) system with two independently controlled excitation sources.
  • Adjusted excitation wavelengths to selectively maximize the excitation of one fluorophore while minimizing the excitation of another.
  • Applied the ME-MPM system to various multi-color imaging applications.

Main Results:

  • Achieved increased signal-to-noise ratios in multi-color imaging.
  • Demonstrated decreased false positive emission bleed-through compared to single-excitation MPM (SE-MPM).
  • Validated the system's effectiveness in diverse multi-color imaging scenarios.

Conclusions:

  • The ME-MPM system is anticipated to increase the adoption of multiphoton microscopy for multi-color applications.
  • ME-MPM serves as a valuable tool for designing and optimizing fluorescent probe pairs for multi-color imaging.
  • This technology enhances the capabilities of advanced imaging techniques like confocal laser scanning microscopy (CLSM).