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Polarization ellipticity compensation in polarization second-harmonic generation microscopy without specimen rotation
Chen-Kuan Chou1, Wei-Liang Chen, Peter Tramyeon Fwu
1National Taiwan University, Department of Physics, No 1 Sec 4 Roosevelt Road, Taipei 106, Taiwan.
Journal of Biomedical Optics
|March 5, 2008
Summary
This study corrects polarization ellipticity in multiphoton microscopy for clearer polarized second-harmonic generation (SHG) imaging of fibrous tissues like muscle and collagen. This method enhances molecular-level structural insights without sample rotation.
Area of Science:
- Biophysics
- Optical Microscopy
- Materials Science
Background:
- Optical microscopy utilizes polarized light to analyze anisotropic samples like collagen and muscle fibers.
- Challenges exist in maintaining polarization control in laser scanning microscopes due to optical component effects.
- Polarization ellipticity alterations can impede accurate molecular information retrieval.
Purpose of the Study:
- To demonstrate a method for correcting polarization ellipticity in multiphoton laser scanning microscopy.
- To enable accurate polarized second-harmonic generation (SHG) imaging of biological tissues.
- To obtain molecular-scale structural information from noncentrosymmetric samples.
Main Methods:
- Employing half-wave and quarter-wave plates to compensate for dichroic mirror-induced polarization ellipticity.
- Utilizing an epi-illuminated multiphoton laser scanning microscope setup.
- Analyzing the excitation polarization dependence of SHG intensity in biological samples.
Main Results:
- Successfully corrected polarization ellipticity alterations caused by the dichroic mirror.
- Acquired polarized SHG images of rat tail tendon and mouse leg muscle.
- Determined the ratio of second-order susceptibility tensor elements for type I collagen and myofibrils.
Conclusions:
- The developed methodology effectively corrects polarization ellipticity for SHG imaging.
- This technique allows for polarized SHG imaging without the need for sample rotation.
- Offers significant potential for detailed molecular and morphological imaging of biological tissues.
