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Multiphoton Fluorescence Excitation

Section Overview:

Multiphoton fluorescence microscopy is a powerful research tool that combines the advanced optical techniques of laser scanning microscopy with long wavelength multiphoton fluorescence excitation to capture high-resolution, three-dimensional images of specimens tagged with highly specific fluorophores.

Review Articles

  • Introduction

    Multiphoton microscopy is a research tool that combines the optical techniques of laser scanning microscopy with long wavelength multiphoton fluorescence excitation to capture three-dimensional images of specimens.

  • Laser Fundamentals

    Explore the fundamentals of lasers and how basic laser systems operate through stimulated emission, and how they are designed to amplify this form of light to create intense and focused beams.

  • Laser Systems for Optical Microscopy

    The lasers employed in optical microscopy are high-intensity monochromatic light sources, which are useful for a variety of techniques including lifetime imaging studies, photobleaching recovery, and TIRFM.

Interactive Java Tutorials

  • Multiphoton Jablonski Diagram

    Two-photon and three-photon excitation occurs as the result of simultaneous fluorophore absorption by photons in a single quantitized event. Explore how fluorescence excitation events occur in multiphoton microscopy.

  • Excitation Region Events

    Examine events occurring in the microscope focal region during specimen excitation using long wavelength visible and near infrared laser illumination in this interactive java tutorial.

  • Excitation Photobleaching Patterns

    Learn more as this interactive java tutorial compares excitation-induced photobleaching patterns that occur near the focal region in both multiphoton and confocal microscopy systems.

  • Ti:Sapphire Mode-Locked Lasers

    Discover and examine the operation of Ti:sapphire lasers over a broad range of near-infrared wavelengths with variable pulse widths and an adjustable applet speed control in this tutorial.

  • Nd:YLF Mode-Locked Pulsed Lasers

    Explore and uncover the operation of a Nd:YLF multi-pass slab laser side-pumped by two collimated diode-laser bars as the tutorial starts off with photons being generated within a Nd:YLF Crystal.

Selected Literature References and Glossary of Terms

  • Selected Literature References

    Listed in this section are periodical location information, book references, as well as providing a listing of selected original research reports from this cutting-edge field of research.

  • Multiphoton Resources on the Web

    Listed in this section are links to resources on the web for multiphoton microscopy including manufacturers, university laboratories, industrial imaging laboratories, technical white papers and tutorials.

Contributing Authors

David W. Piston - Department of Molecular Physiology and Biophysics, Vanderbilt University, 702 Light Hall, Nashville, Tennessee, 37212.

Kenneth R. Spring - Scientific Consultant, Lusby, Maryland, 20657.

John C. Long, Brian O. Flynn, and Michael W. Davidson - National High Magnetic Field Laboratory, 1800 East Paul Dirac Dr., The Florida State University, Tallahassee, Florida, 32310.

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