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Related Experiment Videos

Multiphoton microscopy in life sciences.

K König1

  • 1Laser Microscopy Division, Institute of Anatomy II, Friedrich Schiller University, D-07740 Jena, Germany. kkoe@mti-n.uni-jena.de

Journal of Microscopy
|December 7, 2000
PubMed
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Near infrared (NIR) multiphoton microscopy offers advanced fluorescence imaging and diagnostics in cells and tissues. This technique enables non-invasive optical biopsies and precise intracellular nanosurgery, while also highlighting potential phototoxicity concerns.

Area of Science:

  • Biophysics
  • Optical Engineering
  • Cell Biology

Background:

  • Near-infrared (NIR) multiphoton microscopy is an emerging optical tool for high-resolution imaging in biological systems.
  • It utilizes high-intensity NIR laser radiation for fluorescence excitation, enabling deep tissue penetration and multicolor imaging.

Purpose of the Study:

  • To explore the capabilities of NIR multiphoton microscopy for advanced fluorescence imaging, diagnostics, and nanoprocessing.
  • To investigate its applications in multi-gene detection, non-invasive optical biopsies, and botanical studies.
  • To assess the safety and limitations, including photodamage and phototoxicity, and explore its potential for nanosurgery.

Main Methods:

  • Employing non-resonant two-photon or three-photon excitation with NIR lasers.

Related Experiment Videos

  • Utilizing diffraction-limited focusing to achieve high light intensities (MW cm⁻² to GW cm⁻²).
  • Combining with fluorescence in situ hybridization (FISH) for multi-gene detection.
  • Performing morphological and functional fluorescence imaging of endogenous fluorophores.
  • Investigating cellular responses to high-intensity laser exposure.
  • Utilizing femtosecond NIR lasers for nanosurgical applications.

    • Main Results:

    • Achieved high spatial and temporal resolution fluorescence imaging within living cells and tissues.
    • Enabled multi-gene detection via multiphoton multicolor FISH.
    • Facilitated non-invasive optical biopsies and depth-resolved imaging in botanical samples.
    • Identified mitochondria and Golgi apparatus as major photodamage targets in animal cells, primarily via two-photon excitation.
    • Demonstrated precise intracellular nanosurgery with cut sizes of 100–300 nm using femtosecond NIR lasers, keeping cells viable.

    Conclusions:

    • NIR multiphoton microscopy is a versatile tool for advanced biological imaging, diagnostics, and cellular manipulation.
    • Understanding photodamage mechanisms is crucial for optimizing imaging parameters and ensuring cell viability.
    • The technology holds significant promise for non-invasive diagnostics, fundamental research, and precise intracellular surgery.