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. 2013 Sep 5;4(10):1991-2002.
doi: 10.1364/BOE.4.001991. eCollection 2013.

Precise, motion-free polarization control in Second Harmonic Generation microscopy using a liquid crystal modulator in the infinity space

Chi-Hsiang Lien  1 , Karissa Tilbury, Shean-Jen Chen, Paul J Campagnola
Affiliations

Precise, motion-free polarization control in Second Harmonic Generation microscopy using a liquid crystal modulator in the infinity space

Chi-Hsiang Lien et al. Biomed Opt Express. .

Abstract

Second Harmonic Generation (SHG) microscopy coupled with polarization analysis has great potential for use in tissue characterization, as molecular and supramolecular structural details can be extracted. Such measurements are difficult to perform quickly and accurately. Here we present a new method that uses a liquid crystal modulator (LCM) located in the infinity space of a SHG laser scanning microscope that allows the generation of any desired linear or circular polarization state. As the device contains no moving parts, polarization can be rotated accurately and faster than by manual or motorized control. The performance in terms of polarization purity was validated using Stokes vector polarimetry, and found to have minimal residual polarization ellipticity. SHG polarization imaging characteristics were validated against well-characterized specimens having cylindrical and/or linear symmetries. The LCM has a small footprint and can be implemented easily in any standard microscope and is cost effective relative to other technologies.

Keywords: (110.5405) Polarimetric imaging; (170.6935) Tissue characterization; (180.4315) Nonlinear microscopy; (180.6900) Three-dimensional microscopy; (190.2620) Harmonic generation and mixing.

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Figures

Fig. 1
Fig. 1
Configuration of the SHG microscope with LCM polarization control and polarization analysis.
Fig. 2
Fig. 2
Voltage dependent rotation of the LCM and measured V component of the Stokes vector over the 0-180 degrees of linear rotation.
Fig. 3
Fig. 3
3(a) TPEF linear (left) and circular (right) polarization responses of a GUV labeled with Di-8-ANEPPS. Scale bar = 40 microns. 3(b) The measured TPEF polarization dependent intensity is fit to the theoretical response for a quadratic process.
Fig. 4
Fig. 4
(a) Single SHG optical sections from rat tail tendon. The scale bar is 20 um. (b) SHG intensity as a function of polarization range and the fit to the single axis mode. The error bars are standard deviations.
Fig. 5
Fig. 5
(a) Single SHG optical sections from rat tail skeletal muscle. The scale bar is 20 um. (b) SHG intensity as a function of polarization range and the fit to the single axis mode. The error bars are standard deviations.
Fig. 6
Fig. 6
SHG-CD imaging of optically cleared tendon, showing the RH-CP and LH-CP SHG images and the resulting difference image (SHG-CD). Scale bar = 40 microns.
See this image and copyright information in PMC

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