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Related Experiment Videos

A two-photon and second-harmonic microscope.

Volodymyr Nikolenko1, Boaz Nemet, Rafael Yuste

  • 1Department of Biological Sciences, Columbia University, New York, NY 10027, USA. vn59@columbia.edu

Methods (San Diego, Calif.)
|April 16, 2003
PubMed
Summary
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Custom-built two-photon microscopy offers a cost-effective alternative to commercial systems. This updated design enables advanced imaging and stimulation, making advanced microscopy accessible to more researchers.

Area of Science:

  • Life Sciences
  • Biophysics
  • Microscopy

Background:

  • Two-photon microscopy enables deep tissue imaging with minimal photodamage, crucial for live biological samples.
  • High cost of commercial two-photon microscopes limits accessibility in biological research.
  • Custom-built microscopes offer a potential solution for broader adoption.

Purpose of the Study:

  • To present an updated design for a custom-built two-photon microscope.
  • To demonstrate advanced capabilities including spatiotemporal laser modulation and combined optical techniques.
  • To provide a cost-effective alternative to commercial two-photon microscopy systems.

Main Methods:

  • Modification of an Olympus FluoView laser scanning confocal microscope.
  • Integration of a Pockels cell for arbitrary spatiotemporal laser intensity modulation.

Related Experiment Videos

  • Software control of scanning for precise laser pattern delivery.
  • Combination with second harmonic generation (SHG) measurements.

    • Main Results:

    • Successful implementation of a custom-built two-photon microscope with enhanced control.
    • Demonstration of simultaneous optical imaging, optical stimulation, and SHG measurements.
    • The updated instrument provides a viable, lower-cost alternative to commercial systems.

    Conclusions:

    • Custom-built two-photon microscopy is a feasible and powerful approach for biological research.
    • The developed instrument expands the capabilities of laser scanning microscopes for advanced applications.
    • Increased accessibility of two-photon microscopy can accelerate discoveries in life sciences.