Polarization distortion effects in polarimetric two-photon microscopy.
We present a global analysis of experimental factors affecting polarization responses in two-photon inverted microscopy. The role of reflection optics and high numerical aperture focusing is investigated in two-photon fluorescence, which can be extended to other nonlinear processes. We show that both effects strongly distort polarization responses and can lead to misleading extraction of molecular order information from polarimetric measurements. We describe a model accounting for these effects and develop a calibration technique for the determination of polarization parameters in the sample plane using two-photon fluorescence polarimetry in liquids.
Schön
Peter
P
Institut FRESNEL, MOSAIC, Domaine Universitaire St Jérôme, 13397 Marseille, France.
Munhoz
Fabiana
F
Gasecka
Alicja
A
Brustlein
Sophie
S
Brasselet
Sophie
S
D016477:Artifacts; D007089:Image Enhancement / Q000379:methods*; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D008859:Microscopy, Polarization / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity