Standard Article

Two-photon Microscopy and Imaging

Cell Biology

  1. Patrick Theer,
  2. Bernd Kuhn,
  3. Dorine Keusters,
  4. Winfried Denk

Published Online: 15 SEP 2006

DOI: 10.1002/3527600906.mcb.200500019

Reviews in Cell Biology and Molecular Medicine

Reviews in Cell Biology and Molecular Medicine

How to Cite

Theer, P., Kuhn, B., Keusters, D. and Denk, W. 2006. Two-photon Microscopy and Imaging. Reviews in Cell Biology and Molecular Medicine. .

Author Information

  1. Max Planck Institute for Medical Research, Heidelberg, Germany

Publication History

  1. Published Online: 15 SEP 2006


Multiphoton microscopy (MPM) is an imaging technique that employs signals, such as fluorescence, generated by a multiphoton absorption (MPA) process as its contrast mechanism. Compared to other fluorescence microscopy techniques, MPM often has a higher sensitivity, superior penetration into scattering tissue, and causes less photodamage to the sample and less photobleaching of the fluorophore. These advantages make MPM the method of choice for optophysiological experiments and also make MPM the only technique available for high-resolution in vivo imaging deep within scattering tissue, such as the brain. The two-photon microscope (2PM) is the kind of MPM used in the overwhelming majority of applications since it has virtually all of the specific advantages of MPM. Recent advances in staining techniques including the booming development of genetically encoded fluorophores keep increasing the field of actual and possible MPM applications.


  • Confocal Microscopy (CM);
  • Fluorescence;
  • Laser-scanning Microscopy;
  • Multiphoton Absorption (MPA);
  • Multiphoton Microscopy (MPM);
  • Two-photon Microscopy (2PM);
  • Whole-field Detection;
  • Wide-field Imaging