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Int. J. Mol. Sci. 2013, 14(9), 18520-18534; doi:10.3390/ijms140918520
Article

Differential Polarization Nonlinear Optical Microscopy with Adaptive Optics Controlled Multiplexed Beams

1,2,†
, 1,2,†
, 2
, 1,2
 and 1,2,*
Received: 16 July 2013; in revised form: 21 August 2013 / Accepted: 30 August 2013 / Published: 9 September 2013
(This article belongs to the Special Issue Frontiers of Micro-Spectroscopy in Biological Applications)
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Abstract: Differential polarization nonlinear optical microscopy has the potential to become an indispensable tool for structural investigations of ordered biological assemblies and microcrystalline aggregates. Their microscopic organization can be probed through fast and sensitive measurements of nonlinear optical signal anisotropy, which can be achieved with microscopic spatial resolution by using time-multiplexed pulsed laser beams with perpendicular polarization orientations and photon-counting detection electronics for signal demultiplexing. In addition, deformable membrane mirrors can be used to correct for optical aberrations in the microscope and simultaneously optimize beam overlap using a genetic algorithm. The beam overlap can be achieved with better accuracy than diffraction limited point-spread function, which allows to perform polarization-resolved measurements on the pixel-by-pixel basis. We describe a newly developed differential polarization microscope and present applications of the differential microscopy technique for structural studies of collagen and cellulose. Both, second harmonic generation, and fluorescence-detected nonlinear absorption anisotropy are used in these investigations. It is shown that the orientation and structural properties of the fibers in biological tissue can be deduced and that the orientation of fluorescent molecules (Congo Red), which label the fibers, can be determined. Differential polarization microscopy sidesteps common issues such as photobleaching and sample movement. Due to tens of megahertz alternating polarization of excitation pulses fast data acquisition can be conveniently applied to measure changes in the nonlinear signal anisotropy in dynamically changing in vivo structures.
Keywords: differential microscopy; nonlinear microscopy; multiharmonic generation; genetic algorithms; adaptive optics differential microscopy; nonlinear microscopy; multiharmonic generation; genetic algorithms; adaptive optics
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Samim, M.; Sandkuijl, D.; Tretyakov, I.; Cisek, R.; Barzda, V. Differential Polarization Nonlinear Optical Microscopy with Adaptive Optics Controlled Multiplexed Beams. Int. J. Mol. Sci. 2013, 14, 18520-18534.

AMA Style

Samim M, Sandkuijl D, Tretyakov I, Cisek R, Barzda V. Differential Polarization Nonlinear Optical Microscopy with Adaptive Optics Controlled Multiplexed Beams. International Journal of Molecular Sciences. 2013; 14(9):18520-18534.

Chicago/Turabian Style

Samim, Masood; Sandkuijl, Daaf; Tretyakov, Ian; Cisek, Richard; Barzda, Virginijus. 2013. "Differential Polarization Nonlinear Optical Microscopy with Adaptive Optics Controlled Multiplexed Beams." Int. J. Mol. Sci. 14, no. 9: 18520-18534.


Int. J. Mol. Sci. EISSN 1422-0067 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert