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Proceedings Paper

A method to compensate for the underestimation of collagen with polarized picrosirius red imaging in human artery atherosclerotic plaques
Author(s): C. A. Greiner; S. J. Grainger; J. L. Su; S. P. Madden; J. E. Muller
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Paper Abstract

Although picrosirius red (PSR) is known to be in quantifying collagen under polarized light (PL), commonly used linearly PL can result in an underestimation of collagen, as some of the fibers may appear dark if aligned with the transmission axis of the polarizers. To address this, a sample may be imaged with circularly polarized light at the expense of higher background intensity. However, the quality and alignment of the microscope illumination optics, polarizers and waveplates can still produce imaging variability with circular polarization. A simpler technique was tested that minimized variability and background intensity with linear polarization by acquiring images at multiple angles of histology slide rotation to create a composite co-registered image, permitting the optimal semi-quantitative visualization of collagen.

Linear polarization imaging was performed on PSR stained artery sections. By rotating the slide at 60° intervals while maintaining illumination, polarization and exposure parameters, 6 images were acquired for each section. A composite image was created from the 6 co-registered images, and comprised of the maximum pixel intensity at each point.

Images from any of the 6 rotation positions consistently showed variation in PSR signal. A composite image compensates for this variability, without loss of spatial resolution. Additionally, grayscale analysis showed an increased intensity range of 15 – 50% with a linearly polarized composite image over a circularly polarized image after background correction, indicating better SNR. This proposed technique will be applied in the development of a near infrared spectroscopy algorithm to detect vulnerable atherosclerotic plaques in vivo.

Paper Details

Date Published: 6 April 2016
PDF: 5 pages
Proc. SPIE 9711, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues IX, 97111R (6 April 2016); doi: 10.1117/12.2211388
Show Author Affiliations
C. A. Greiner, Infraredx, Inc. (United States)
S. J. Grainger, Infraredx, Inc. (United States)
J. L. Su, Infraredx, Inc. (United States)
S. P. Madden, Infraredx, Inc. (United States)
J. E. Muller, Ctr. for Integration of Medicine and Innovative Technology (United States)

Published in SPIE Proceedings Vol. 9711:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues IX
Daniel L. Farkas; Dan V. Nicolau; Robert C. Leif, Editor(s)

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