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

TL;DR

This study introduces a quantitative polarized light microscopy method to diagnose amyloid fibrils in tissue samples without using Congo red dye, improving accuracy and simplicity over traditional qualitative techniques.

Contribution

The paper presents an improved, dye-free PLM technique that quantitatively detects amyloid fibrils using Fourier analysis, reducing misdiagnosis and technical limitations of conventional methods.

Findings

Quantitative birefringence analysis correlates with amyloid density.

Method distinguishes amyloid from other tissue structures.

No need for Congo red staining simplifies diagnosis.

Abstract

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.