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目的探讨组织化学染色技术是否可以应用于塑化标本并验证染色塑化标本是否具有自发荧光。方法选取1个手掌标本经超薄生物塑化技术做成组织块,进行连续切片,切片数量56张,并按切片顺序进行染色处理:原始切片,苏木素-伊红染色,凡尔霍夫-酸性品红染色,亚甲蓝-天青Ⅱ号染色,在扫描仪、光学显微镜和激光扫描共焦显微镜下观察染色效果和组织结构特点并进行比较。结果 3种染色技术都可应用于塑化切片染色。苏木素-伊红染色显示肌肉、结缔组织呈红色或紫红色,骨质呈紫蓝色或蓝色。凡尔霍夫-酸性品红染色显示弹力纤维呈黑色,胶原呈红色,其他组织为黄色或棕黄色。亚甲蓝-天青Ⅱ号染色显示肌腱呈紫红色,骨质呈粉红色,软骨呈紫罗兰,其他组织呈紫色。但细胞内结构的染色效果并不理想。在激光扫描共焦显微镜下,胶原纤维、弹力纤维和肌肉纤维具有自发荧光,结构清晰可辨。结论常用的组织化学染色技术适合于超薄塑化切片染色,染色切片的各种组织结构比未染色的观察效果好。染色后,塑化切片中具有自发荧光的结构在激光扫描共焦显微镜下亦可清晰显影。
Objective To investigate whether histochemical staining technique can be used to plasticize specimens and verify whether the stained plasticized specimens have autofluorescence. Methods One palm specimen was made into tissue blocks by ultrathin bioplastics technology. The number of slices was 56 and the number of slices was 56. The original slices, hematoxylin and eosin staining, Magenta staining, methylene blue - Azure II staining, the scanning electron microscopy and laser scanning confocal microscope to observe the dyeing effect and the characteristics of tissue structure and comparison. Results All three staining techniques were applied to plasticized sections. Hematoxylin - eosin staining showed muscle, connective tissue was red or purple, bone was purple blue or blue. Verhoeff-Acid magenta staining showed elastic fibers in black, collagen in red, and other tissues in yellow or brown. Methylene blue - Azure II staining showed a reddish tendon, pink bone, cartilage was violets, other tissues were purple. However, the intracellular structure of the staining effect is not satisfactory. Under laser scanning confocal microscope, collagen fibers, elastic fibers and muscle fibers with autofluorescence, the structure is clearly identifiable. Conclusion The commonly used techniques of histochemical staining are suitable for ultrathin plasticized sections dyeing. The histological structure of the dyed sections is better than the undyed ones. After staining, the structure with autofluorescence in plasticized sections can also be clearly developed under a laser scanning confocal microscope.