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

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.

Multiphoton autofluorescence and second-harmonic generation imaging of the tooth.

In this study, we used an epi-illuminated multiphoton microscope to image three main components of ex vivo human tooth. In particular, we obtained two-photon excited autofluorescence (AF) and second-harmonic generation (SHG) images of the enamel, dentin, and periodontal ligaments (PLs) and constructed three-dimensional projections of sequentially and axially acquired images. We found that the enamel has a strong two-photon AF signal, clearly revealing the structures of the enamel rods.