Observations of amyloid breakdown by proteases over time using scanning acoustic microscopy

Abstract

Amyloid consists of insoluble beta-fibrillar proteins with stable structures. The Congo red staining method for histologically detecting amyloid is unsuitable for quantitatively assessing amyloid fibers. Scanning acoustic microscopy (SAM) detects the attenuation of sound (AOS) through sections. This study aimed to clarify whether AOS values reflected the amount of amyloid fibril degradation in tissues. Formalin-fixed paraffin-embedded unstained sections of various types of amyloidosis were digested with different endopeptidases. The AOS images after digestion were observed over time via SAM. The corresponding Congo red-stained images were followed to identify the amyloid. The amyloid and nonamyloid portions were statistically examined over time to determine the changes in the AOS values. Most of the amyloid areas showed significantly different AOS values from nonamyloid portions before digestion and significantly decreased after digestion; these findings corresponded with the disappearance and waning of the Congo red staining in the light microscopic images. Some nonamyloid areas with high AOS masked the reduction in AOS in the amyloid areas. The method used in this study may help detect the amyloid quantity and determine the appropriate treatment method for removing amyloid deposits from tissues.