On the fundamental imaging-depth limit in two-photon microscopy

J Opt Soc Am A Opt Image Sci Vis. 2006 Dec;23(12):3139-49. doi: 10.1364/josaa.23.003139.


We have analyzed how the maximal imaging depth of two-photon microscopy in scattering samples depends on properties of the sample and the imaging system. We find that the imaging depth increases with increasing numerical aperture and staining inhomogeneity and with decreasing excitation-pulse duration and scattering anisotropy factor, but is ultimately limited by near-surface fluorescence with slight improvements possible using special detection strategies.

MeSH terms

  • Algorithms*
  • Image Enhancement / methods*
  • Image Interpretation, Computer-Assisted / methods*
  • Imaging, Three-Dimensional / methods*
  • Information Storage and Retrieval / methods*
  • Microscopy, Fluorescence, Multiphoton / instrumentation
  • Microscopy, Fluorescence, Multiphoton / methods*
  • Phantoms, Imaging
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Signal Processing, Computer-Assisted*