Miura Katsutoshi, Iwashita Toshihide
Department of Regenerative and Infectious Pathology, Hamamatsu University School of Medicine, 1-20-1 Handa-yama, Chuoku, Hamamatsu, Shizuoka, 431-3192, Japan.
Department of Health Science, Hamamatsu University School of Medicine, 1-20-1 Handa-yama, Chuoku, Hamamatsu, Shizuoka, 431-3192, Japan.
Sci Rep. 2025 Aug 11;15(1):29300. doi: 10.1038/s41598-025-15598-1.
Detecting specific tissue components is valuable in histology. Scanning acoustic microscopy (SAM) measures the attenuation-of-sound (AOS) through tissue sections, enabling the generation of histological images without staining. AOS values decrease as tissues degrade. In this study, we enzymatically digested target components and monitored the process using AOS imaging over time. Additionally, we applied specific dyes and antibodies to inhibit enzyme activity and preserve target component. Collagenase digested the bone to clearly visualise the internal structure. The target component showed a distinct decline in AOS values. Actinase digested the artery except for amyloid deposits, which were detected by Congo red staining. Actinase-digested lymphoid cells remained positive for horseradish peroxidase (HRP) staining. Amylase digested some corpora amylacea (CA) in the brain, which became negative for periodic acid-Schiff (PAS) staining and diminished in size under electron microscopy. DNase digested and deleted cell nuclei, except for those stained with HRP. Residual nuclear images of AOS matched those of light microscopy. Enzyme-specific inhibition of enzymes preserved the target cells and materials. Our method offers a practical method for intentionally deleting or retaining target components in a section. Furthermore, it provides a means to adjust and compare the degree of degradation using AOS values.
在组织学中，检测特定的组织成分具有重要价值。扫描声学显微镜（SAM）通过组织切片测量声音衰减（AOS），能够生成无需染色的组织学图像。随着组织降解，AOS值会降低。在本研究中，我们用酶消化目标成分，并通过AOS成像随时间监测该过程。此外，我们应用特定的染料和抗体来抑制酶活性并保存目标成分。胶原酶消化骨骼以清晰显示其内部结构。目标成分的AOS值出现明显下降。肌动蛋白酶消化动脉，除淀粉样沉积物外，刚果红染色可检测到这些沉积物。经肌动蛋白酶消化的淋巴细胞对辣根过氧化物酶（HRP）染色仍呈阳性。淀粉酶消化大脑中的一些淀粉样体（CA），这些淀粉样体对过碘酸希夫（PAS）染色呈阴性，并且在电子显微镜下尺寸减小。脱氧核糖核酸酶消化并去除细胞核，除了那些用HRP染色的细胞核。AOS的残留核图像与光学显微镜的图像相匹配。对酶进行酶特异性抑制可保存目标细胞和物质。我们的方法提供了一种在切片中有意删除或保留目标成分的实用方法。此外，它提供了一种利用AOS值来调整和比较降解程度的手段。
