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Research Papers

Polarization ellipticity compensation in polarization second-harmonic generation microscopy without specimen rotation

[+] Author Affiliations
Chen-Kuan Chou, Wei-Liang Chen, Peter Tramyeon Fwu

National Taiwan University, Department of Physics, No 1 Sec 4 Roosevelt Road, Taipei 106, Taiwan

Sung-Jan Lin

National Taiwan University, Institute of Biomedical Engineering, No 1 Sec 4 Roosevelt Road, Taipei 106, Taiwan

Hsuan-Shu Lee

National Taiwan University Hospital, Department of Internal Medicine, No 7 Chung San South Road, Taipei 100, Taiwan

Chen-Yuan Dong

National Taiwan University, Department of Physics, No 1 Sec 4 Roosevelt Road, Taipei 106, Taiwan

J. Biomed. Opt. 13(1), 014005 (June 22, 2007October 09, 2007October 09, 2007January 02, 2008). doi:10.1117/1.2824379
History: Received June 22, 2007; Revised October 09, 2007; Accepted October 09, 2007; Published January 02, 2008
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In imaging anisotropic samples with optical microscopy, a controlled, polarized light source can be used to gain molecular information of fibrous materials such as muscles and collagen fibers. However, the delivery of the polarized excitation light source in a system such as a laser scanning optical microscope often encounters the problem of the polarization ellipticity altering effects of the optical components. Using a half-wave plate and a quarter-wave plate, we demonstrate that the polarization ellipticity altering effect of the dichroic mirror in an epi-illuminated multiphoton laser scanning microscope can be corrected, and that this approach can be used to obtain polarized second-harmonic generation (SHG) images of rat tail tendon and mouse leg muscle. The excitation polarization dependence of the SHG intensity is fitted to determine the ratio of the second-order susceptibility tensor elements associated with type I collagen in the rat tail tendon and myofibril in the mouse leg muscle. Our methodology can be applied to polarized SHG imaging without sample rotation. This approach has great potential for imaging noncentrosymmetric biological samples, providing structural information on the molecular scale in addition to morphological information of tissues.

Figures in this Article
© 2008 Society of Photo-Optical Instrumentation Engineers

Citation

Chen-Kuan Chou ; Sung-Jan Lin ; Hsuan-Shu Lee ; Chen-Yuan Dong ; Wei-Liang Chen, et al.
"Polarization ellipticity compensation in polarization second-harmonic generation microscopy without specimen rotation", J. Biomed. Opt. 13(1), 014005 (June 22, 2007October 09, 2007October 09, 2007January 02, 2008). ; http://dx.doi.org/10.1117/1.2824379


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