Maintaining polarization in polarimetric multiphoton microscopy

J Biophotonics. 2015 Nov;8(11-12):884-8. doi: 10.1002/jbio.201400116. Epub 2015 Feb 17.


Polarimetric measurements in multiphoton microscopy can reveal information about the local molecular order of a sample. However, the presence of a dichroic through which the excitation beam propagates will generally scramble its polarization. We propose a simple scheme whereby a second properly-oriented compensation dichroic is used to negate any alteration regardless of the wavelength and the initial polarization. We demonstrate how this robust and rapid approach simplifies polarimetric measurements in second-harmonic generation, two-photon excited fluorescence and coherent anti-Stokes Raman scattering. Illustration of the polarization maintaining strategy with the compensating dichroic oriented such that its s- and p-axes are interchanged with these of the primary dichroic.

Keywords: multiphoton microscopy; neurophotonic; neuroscience; polarimetric microscopy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Collagen / chemistry
  • Demyelinating Diseases / pathology
  • Equipment Design
  • Fluorescent Dyes
  • Lysophosphatidylcholines
  • Microscopy, Polarization / instrumentation
  • Microscopy, Polarization / methods*
  • Models, Theoretical
  • Myelin Sheath / chemistry
  • Myelin Sheath / pathology
  • Oxazines


  • Fluorescent Dyes
  • Lysophosphatidylcholines
  • Oxazines
  • Collagen
  • nile red