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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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Updated: Mar 17, 2026

Low-cost Custom Fabrication and Mode-locked Operation of an All-normal-dispersion Femtosecond Fiber Laser for Multiphoton Microscopy
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Two-photon microscopy using fiber-based nanosecond excitation.

Sebastian Karpf1, Matthias Eibl2, Benjamin Sauer2

  • 1Lehrstuhl für BioMolekulare Optik, Fakultät für Physik, Ludwig-Maximilians-Universität München, Oettingenstr. 67, 80538 Munich, Germany; Department of Electrical Engineering, University of California, Los Angeles, California 90095, USA.

Biomedical Optics Express
|July 23, 2016
PubMed
Summary

This study presents a novel fiber-based two-photon excitation fluorescence (TPEF) microscope using nanosecond pulses for improved in vivo internal organ imaging. This approach overcomes limitations of femtosecond pulses in endoscopic applications, enabling deeper tissue visualization.

Keywords:
(060.2350) Fiber optics imaging(060.4370) Nonlinear optics, fibers(140.3510) Lasers, fiber(180.2520) Fluorescence microscopy(180.4315) Nonlinear microscopy

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Area of Science:

  • Biomedical Optics
  • Microscopy
  • Medical Imaging

Background:

  • Two-photon excitation fluorescence (TPEF) microscopy offers sensitive deep tissue imaging up to 1000 micrometers.
  • In vivo imaging of internal organs requires endoscopic beam delivery, which is limited by pulse broadening in optical fibers.

Purpose of the Study:

  • To develop an endoscope-compatible TPEF microscope for in vivo internal organ imaging.
  • To overcome the pulse broadening limitations of femtosecond pulses in endoscopic fiber delivery.

Main Methods:

  • Utilized nanosecond pulses at low repetition rates instead of traditional femtosecond pulses.
  • Developed a fiber-based TPEF microscope system for endoscopic applications.
  • Derived and demonstrated the dependence of TPEF signal on the laser source's duty cycle.

Main Results:

  • Nanosecond pulses circumvented linear and non-linear pulse broadening issues in optical fibers.
  • Demonstrated that TPEF signal intensity is proportional to the laser duty cycle at a given continuous wave power.
  • Achieved single-shot two-photon fluorescence lifetime measurements due to higher pulse energy.

Conclusions:

  • The developed nanosecond pulse TPEF microscope is suitable for endoscope-based in vivo imaging.
  • This technique enhances the feasibility of deep tissue imaging in internal organs.
  • Offers potential for advanced diagnostics through fluorescence lifetime measurements.