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Live cell imaging by multifocal multiphoton microscopy.

M Straub1, P Lodemann, P Holroyd

  • 1High Resolution Optical Microscopy Group, Max-Planck-Institute for Biophysical Chemistry, Göttingen, Germany. mstraub@gwdg.de

European Journal of Cell Biology
|November 23, 2000
PubMed
Summary
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Multifocal multiphoton microscopy (MMM) enables rapid, high-resolution 3D live cell imaging. This technique visualizes cellular structures and organelles with exceptional speed and clarity.

Area of Science:

  • Biophysics
  • Cell Biology
  • Microscopy

Background:

  • Multifocal multiphoton microscopy (MMM) offers parallel excitation for faster imaging.
  • It combines optical sectioning and reduced phototoxicity, ideal for live cells.
  • Standard multiphoton microscopy has limitations in speed for dynamic cellular processes.

Purpose of the Study:

  • To demonstrate the application of MMM for high-resolution 3D live cell imaging.
  • To visualize nuclear chromatin and acidic organelles in neuroendocrine and chromaffin cells.
  • To reconstruct cell and neurite morphology in three dimensions.

Main Methods:

  • Utilized multifocal multiphoton microscopy (MMM) for live cell imaging.
  • Employed various stains including acridine orange, FM1-43, Fast DiA, and GFP transfection.

Related Experiment Videos

  • Acquired image stacks rapidly for 3D reconstruction using volume-rendering and isosurface plots.

    • Main Results:

    • Successfully visualized nuclear chromatin and the 3D distribution of acidic organelles in PC12 and bovine chromaffin cells.
    • Monitored neurite extensions in nerve growth factor-differentiated PC12 cells.
    • Generated detailed 3D cell and neurite morphology reconstructions from image stacks acquired in seconds.

    Conclusions:

    • MMM is a technically simple and highly effective method for fast 3D live cell imaging.
    • The technique provides high-resolution visualization of cellular structures.
    • MMM significantly enhances the speed of live cell imaging compared to conventional methods.