Guselnikova V V, Fedorova E A, Gudkova A J, Shavlovsky M M, Korzhevskii D E
Institute of Experimental Medicine.
Pavlov First Saint Petersburg State Medical University.
Ter Arkh. 2022 May 26;94(4):473-478. doi: 10.26442/00403660.2022.04.201464.
To compare efficiency and specific features of transthyretin amyloid staining by different histological dyes and thus to assess their suitability for diagnostic purposes.
Samples of left and right heart ventricles were taken from patients over 70 years-old of both genders (n=10) with immunohistochemically verified transthyretin amyloidosis (ATTR). All samples were stained with Congo red, Alcian blue, toluidine blue and methylene violet.
Specificity and sensitivity of Congo red staining was comparable to those of immunohistochemical staining. For verification of amyloid presence after Congo red staining one could use fluorescent microscopy instead of polarization microscopy. It allows a more accurate diagnosis of amyloidosis. Confocal microscopy with spectral unmixing improves detection sensitivity of amyloid by elimination of background fluorescence of muscle tissue and autofluorescence of lipofuscin. Alcian blue staining gives the same result as Congo red. In addition, its less labor-intensive and free of false-positive and false-negative results caused by final processing of slide preparation. Toluidine blue and methylene violet develop metachromatic staining upon binding to transthyretin fibrils, likely due to specific biochemical features of these fibrils.
The most reliable method for histochemical diagnosis of ATTR is the Congo red staining with subsequent analysis using fluorescence or confocal microscopy. For diagnostic screening, the use of Sodium sulphate-Alcian blue staining method is highly promising. Metachromatic stains are less effective for ATTR diagnosis.
比较不同组织学染料对转甲状腺素蛋白淀粉样变的染色效率和特点,从而评估其用于诊断目的的适用性。
从70岁以上的男女患者(n = 10)中获取左、右心室样本,这些患者经免疫组织化学证实患有转甲状腺素蛋白淀粉样变(ATTR)。所有样本均用刚果红、阿尔辛蓝、甲苯胺蓝和亚甲蓝进行染色。
刚果红染色的特异性和敏感性与免疫组织化学染色相当。刚果红染色后,可用荧光显微镜而非偏振显微镜来验证淀粉样物质的存在。这能更准确地诊断淀粉样变。采用光谱解混的共聚焦显微镜通过消除肌肉组织的背景荧光和脂褐素的自发荧光,提高了淀粉样物质的检测灵敏度。阿尔辛蓝染色结果与刚果红相同。此外,其操作强度较小,且不会因玻片制备的最终处理而产生假阳性和假阴性结果。甲苯胺蓝和亚甲蓝与转甲状腺素蛋白原纤维结合后会产生异染性染色,这可能是由于这些原纤维的特定生化特性所致。
ATTR组织化学诊断最可靠的方法是刚果红染色,随后使用荧光或共聚焦显微镜进行分析。对于诊断筛查,硫酸钠 - 阿尔辛蓝染色法极具前景。异染性染色对ATTR诊断效果较差。