Polarimetry-based method to extract geometry-independent metrics of tissue anisotropy.

Recently, we have used polarimetry as a method for assessing the linear retardance of infarcted myocardium. While linear retardance reflects tissue anisotropy, experimental geometry has a confounding effect due to dependence of the linear retardance on the orientation of the sample with respect to the probing beam. Here, polarimetry imaging of an 8mm diameter birefringent polystyrene sphere of known anisotropy axis was used to test a dual-projection method by which the anisotropy axis and its true magnitude can be reconstructed, thus eliminating the confounding effect of anisotropy axis orientation. Feasibility is demonstrated in ex-vivo tissue imaging.

Two-photon microscopy of healthy, infarcted and stem-cell treated regenerating heart.

Two-photon excitation autofluorescence (produced in myocytes) and second-harmonic generation (produced mainly by collagen) allow label-free visualization of these two important components of myocardium. Because of their different emission wavelengths, these two signals can be separated spectrally. Here, we examine two-photon microscopy images of healthy, infarcted and stem-cell treated rat hearts. We find that in infarcted heart, regions distant from the site of infarct are similar to healthy tissue in composition (mostly myocytes, very little collagen) and organization (densely packed myocytes), but infarct regions are characterized by sparse myocytes and high collagen content indicative of scar tissue formation. Stem cell treated hearts, in contrast, show regions of intertwined myocytes and collagen throughout the infarct, suggesting reduced tissue damage. Finally, these results offer interesting insights into our ongoing polarized light studies of cardiac tissue anisotropy, and reveal that both tissue composition and tissue micro-organization are reflected in polarization-measured linear retardance values.

Polarization birefringence measurements for characterizing the myocardium, including healthy, infarcted, and stem-cell-regenerated tissues.

Myocardial infarction leads to structural remodeling of the myocardium, in particular to the loss of cardiomyocytes due to necrosis and an increase in collagen with scar formation. Stem cell regenerative treatments have been shown to alter this remodeling process, resulting in improved cardiac function. As healthy myocardial tissue is highly fibrous and anisotropic, it exhibits optical linear birefringence due to the different refractive indices parallel and perpendicular to the fibers. Accordingly, changes in myocardial structure associated with infarction and treatment-induced remodeling will alter the anisotropy exhibited by the tissue. Polarization-based linear birefringence is measured on the myocardium of adult rat hearts after myocardial infarction and compared with hearts that had received mesenchymal stem cell treatment. Both point measurement and imaging data show a decrease in birefringence in the region of infarction, with a partial rebound back toward the healthy values following regenerative treatment with stem cells. These results demonstrate the ability of optical polarimetry to characterize the micro-organizational state of the myocardium via its measured anisotropy, and the potential of this approach for monitoring regenerative treatments of myocardial infarction.