Li Bo, Wu Ying, Gao Xiao-Ming
Department of Immunology, Peking University Health Science Center, Peking University, Beijing 100083, China.
Histochem J. 2002 Jun-Jul;34(6-7):299-303. doi: 10.1023/a:1023325213198.
Pyronin Y has long been used, in combination with other dyes such as Methyl Green, as a differential stain for nucleic acids in paraffin tissue sections. It also forms fluorescent complexes with double-stranded nucleic acids, especially RNA, enabling semi-quantitative analysis of cellular RNA in flow cytometry. However, the possibility of using pyronin Y as a fluorescent stain for paraffin tissue sections has rarely been investigated. We herein report that in sections stained with Methyl Green-pyronin Y, red blood cells, elastic fibre of blood vessels, zymogen granules of pancreatic acinar cells, surface membrane of heptocytes and kidney tubular cells showed strikingly strong green and/or red fluorescence, while the nuclei of cells appeared non-fluorescent. The use of confocal laser-scanning microscope greatly improved the resolution and selectivity of the fluorescent images. Staining with pyronin Y alone gave similar results in terms of fluorescence properties of the specimens. Pretreatment of paraffin sections with RNase significantly reduced cytoplasmic pyronin Y staining as judged by transmission light microscopy, but it had little effect on the fluorescence intensity of red blood cells, elastic fibres and zymogen granules.
派洛宁Y长期以来一直与甲基绿等其他染料联合使用，作为石蜡组织切片中核酸的鉴别染色剂。它还能与双链核酸，尤其是RNA形成荧光复合物，从而在流式细胞术中对细胞RNA进行半定量分析。然而，将派洛宁Y用作石蜡组织切片荧光染色剂的可能性鲜有研究。我们在此报告，在用甲基绿-派洛宁Y染色的切片中，红细胞、血管弹性纤维、胰腺腺泡细胞的酶原颗粒、肝细胞表面膜和肾小管细胞呈现出极强的绿色和/或红色荧光，而细胞核则无荧光。共聚焦激光扫描显微镜的使用极大地提高了荧光图像的分辨率和选择性。单独用派洛宁Y染色在标本的荧光特性方面得到了类似的结果。用核糖核酸酶对石蜡切片进行预处理，通过透射光显微镜判断，可显著降低细胞质中派洛宁Y的染色，但对红细胞、弹性纤维和酶原颗粒的荧光强度影响不大。
