Space multiplexed multiphoton microscopy by temporal focusing of ultrashort pulses

Dan Oron, Eran Tal, Yaron Silberberg

Research output: Contribution to journalConference articlepeer-review


The ability to perform optical sectioning is one of the great advantages of laser-scanning microscopy. This introduces, however, a number of difficulties due to the scanning process, such as lower frame rates due to the serial acquisition process. Here we show that by introducing spatiotemporal pulse shaping techniques to multiphoton microscopy it is possible to obtain full-frame depth resolved imaging completely without scanning. Our method relies on temporal focusing of the illumination pulse. The pulsed excitation field is compressed as it propagates through the sample, reaching its shortest duration at the focal plane, before stretching again beyond it. Combining temporal focusing with line-scanning microscopy results in an enhanced depth resolution, equivalent to that achieved by point scanning. Both the scanningless and the line-scanning techniques are applied to obtain depth-resolved two-photon excitation fluorescence (TPEF) images of drosophila egg-chambers.

Original languageEnglish
Article number10
Pages (from-to)35-43
Number of pages9
JournalProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Publication statusPublished - Jul 21 2005
EventMultiphoton Microscopy in the Biomedical Sciences V - San Jose, CA, United States
Duration: Jan 23 2005Jan 25 2005


  • Laser-scanning
  • Microscopy
  • Nonlinear optics
  • Two-photon excitation fluorescence

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Biomaterials
  • Atomic and Molecular Physics, and Optics
  • Radiology Nuclear Medicine and imaging

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