Optimal repetition rate and pulse duration studies for two photon imaging

Shamil Mirkhanov, Adrian H. Quarterman, Connor J.C.P. Smyth, Bavishna B. Praveen, Paul Appleton, Calum Thomson, Samuel Swift, Keith G. Wilcox

Research output: Chapter in Book/Report/Conference proceedingConference contribution

2 Citations (Scopus)


Multiphoton imaging (MPI) is an important fluorescence microscopy technique that allows deep tissue and in-vivo imaging with high selectivity. According to theory, two-photon signal is proportional to the product of the peak power and the average power, allowing optimization of key imaging parameters of the excitation laser, such as average power, repetition rate and pulse duration. Recent progress in compact ultrafast lasers including femtosecond fiber lasers and optically pumped semiconductor lasers makes direct control of these parameters possible. In order to investigate the optimum laser parameters for two photon imaging we experimentally study the effects of repetition rate between 2.85 and 90 MHz and pulse duration between 336 fs and 3.5 ps on two photon signal in SYTOX Green labeled mouse intestine sections at 1030 nm. We found that the optimum repetition rate for this sample is in the range 20 - 40 MHz, depending on average power, and that the pulse duration has no effect on the MPI signal provided that the average power can be adjusted to keep the product of average and peak power constant.

Original languageEnglish
Title of host publicationMultiphoton Microscopy in the Biomedical Sciences XVII
PublisherSPIE-International Society for Optical Engineering
ISBN (Electronic)9781510605794
Publication statusPublished - 28 Jan 2017
EventSPIE Photonics West 2017: Multiphoton Microscopy in the Biomedical Sciences XVII - Moscone Center, San Francisco, United States
Duration: 29 Jan 201731 Jan 2017
http://spie.org/conferences-and-exhibitions/past-conferences-and-exhibitions/photonics-west-2017 (Link to Conference details)


ConferenceSPIE Photonics West 2017
Country/TerritoryUnited States
CitySan Francisco
Internet address


  • fiber lasers
  • fluorescence microscopy
  • Multiphoton microscopy
  • ultrafast lasers

ASJC Scopus subject areas

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


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