Polarization modulation laser scanning microscopy: A powerful tool to image molecular orientation and order

Abstract

To image the orientational order in a broad class of biological and manufactured materials, a new microscope has been developed that integrates laser scanning microscopy with polarization modulation polarimetry. Polarimetry allows quantitative characterization of the molecular orientation and the degree of order through characterization of optical anisotropy. Combined with laser scanning microscopy, it is used here to image the anisotropy with high spatial resolution, sensitivity, and speed. The design of the microscope is presented; and the vast improvement in sensitivity achieved using PM-LSM over conventional polarization microscopy is illustrated by imaging the linear dichroism of ultrathin Langmuir–Blodgett polymer films. PM-LSM allows imaging of the magnitude and orientation of linear dichroism in films as thin as three molecular layers (~66 Å) at high resolution by rastering a diffraction limited spot of laser light across the sample. The rate of image acquisition is over 2000 pixels/s, two to three orders of magnitude faster than the previous methods of imaging optical anisotropy.

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