Deep tissue multiphoton microscopy using longer wavelength excitation

Opt Express. 2009 Aug 3;17(16):13354-64. doi: 10.1364/oe.17.013354.


We compare the maximal two-photon fluorescence microscopy (TPM) imaging depth achieved with 775-nm excitation to that achieved with 1280-nm excitation through in vivo and ex vivo TPM of fluorescently-labeled blood vessels in mouse brain. We achieved high contrast imaging of blood vessels at approximately twice the depth with 1280-nm excitation as with 775-nm excitation. An imaging depth of 1 mm can be achieved in in vivo imaging of adult mouse brains at 1280 nm with approximately 1-nJ pulse energy at the sample surface. Blood flow speed measurements at a depth of 900 mum are performed.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Computer Simulation
  • Computer-Aided Design
  • Equipment Design
  • Equipment Failure Analysis
  • Image Enhancement / instrumentation*
  • Light
  • Lighting / instrumentation*
  • Microscopy, Fluorescence, Multiphoton / instrumentation*
  • Models, Theoretical
  • Scattering, Radiation
  • Sensitivity and Specificity