Quantitative diagnosis of amyloid without Congo red staining using polarized light microscopy

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Main Authors: Lailey, Owen, Alais, Maria Agustina, Wang, Liuhe, Chahal, Pinki, Cory, David G., Khoo, Timothy, Olkhov-Mitsel, Ekaterina, Sarenac, Dusan, Pushin, Dmitry A., Mirkovic, Jelena
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Published: 2025
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author Lailey, Owen
Alais, Maria Agustina
Wang, Liuhe
Chahal, Pinki
Cory, David G.
Khoo, Timothy
Olkhov-Mitsel, Ekaterina
Sarenac, Dusan
Pushin, Dmitry A.
Mirkovic, Jelena
author_facet Lailey, Owen
Alais, Maria Agustina
Wang, Liuhe
Chahal, Pinki
Cory, David G.
Khoo, Timothy
Olkhov-Mitsel, Ekaterina
Sarenac, Dusan
Pushin, Dmitry A.
Mirkovic, Jelena
contents Amyloidosis is a protein misfolding disease caused by the deposition of large, insoluble aggregates (amyloid fibrils) of protein in a tissue, which has been associated with various conditions, such as lymphoid disorders, Alzheimer's disease, diabetes mellitus type 2, chronic inflammatory processes, and cancers. Amyloid fibrils are commonly diagnosed by qualitative observation of green birefringence from Congo red stained biopsy tissue samples under polarized light, a technique that is limited by lack of specificity, dependence on subjective interpretation, and technical constraints. Studies emphasize the utility of quantitative polarized light microscopy (PLM) methodology to diagnose amyloid fibrils in Congo red stained tissues. However, while Congo red enhances the intrinsic birefringence of amyloid fibrillar structures, there are significant disadvantages such as the appearance of multiple non-green colors under polarized light and binding to other structures, which may result in misdiagnoses with Congo red dye and inconclusive explanations. In this work, we present an improved PLM methodology for quantitative detection of amyloid fibrils without requiring Congo red staining. We perform PLM measurements on four tissues: abdominal subcutaneous tissue biopsy, duodenal biopsy, thyroid biopsy, and breast biopsy, both with Congo red stain and H\&E stain, and through Fourier analysis quantify birefringence, birefringent axis orientation, dichroism, optical activity, and relative amyloid density. These results emphasize a quantitative analysis for amyloid diagnosis rooted in Fourier signal harmonics that does not require Congo red dye and paves the way for rapid, simple, and accurate diagnosis of amyloid fibrils.
format Preprint
id arxiv_https___arxiv_org_abs_2502_12349
institution arXiv
publishDate 2025
record_format arxiv
spellingShingle Quantitative diagnosis of amyloid without Congo red staining using polarized light microscopy
Lailey, Owen
Alais, Maria Agustina
Wang, Liuhe
Chahal, Pinki
Cory, David G.
Khoo, Timothy
Olkhov-Mitsel, Ekaterina
Sarenac, Dusan
Pushin, Dmitry A.
Mirkovic, Jelena
Medical Physics
Amyloidosis is a protein misfolding disease caused by the deposition of large, insoluble aggregates (amyloid fibrils) of protein in a tissue, which has been associated with various conditions, such as lymphoid disorders, Alzheimer's disease, diabetes mellitus type 2, chronic inflammatory processes, and cancers. Amyloid fibrils are commonly diagnosed by qualitative observation of green birefringence from Congo red stained biopsy tissue samples under polarized light, a technique that is limited by lack of specificity, dependence on subjective interpretation, and technical constraints. Studies emphasize the utility of quantitative polarized light microscopy (PLM) methodology to diagnose amyloid fibrils in Congo red stained tissues. However, while Congo red enhances the intrinsic birefringence of amyloid fibrillar structures, there are significant disadvantages such as the appearance of multiple non-green colors under polarized light and binding to other structures, which may result in misdiagnoses with Congo red dye and inconclusive explanations. In this work, we present an improved PLM methodology for quantitative detection of amyloid fibrils without requiring Congo red staining. We perform PLM measurements on four tissues: abdominal subcutaneous tissue biopsy, duodenal biopsy, thyroid biopsy, and breast biopsy, both with Congo red stain and H\&E stain, and through Fourier analysis quantify birefringence, birefringent axis orientation, dichroism, optical activity, and relative amyloid density. These results emphasize a quantitative analysis for amyloid diagnosis rooted in Fourier signal harmonics that does not require Congo red dye and paves the way for rapid, simple, and accurate diagnosis of amyloid fibrils.
title Quantitative diagnosis of amyloid without Congo red staining using polarized light microscopy
topic Medical Physics
url https://arxiv.org/abs/2502.12349