组织化学染色技术在超薄生物塑化标本中的应用
Histochemical staining techniques applied in ultrathin sheet plastination slice
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 and what staining techniques are applied to the ultrathin sheet plastination slice and whether the stained specimen is of autofluorescences. Methods A cadaveric hand block was plastinated and then sectioned as a series of 300-400 μm thick transverse sections. A total of 56 slices in total. Alternative sections were stained with hematoxylin -eosin staining (HE), Verhoeff -Van Gieson staining(VVG) or methylene blue and azureⅡstaining(MA). The stained slices were examined under a light microscope and a confocal microscope. Results The plastinated slices were stained with the three staining methods. HE staining revealed the muscle and connective tissues were red or violet, bone was violet or blue; VVG staining showed the elastic fibers was black, the collagen was red, and other tissues were yellow. MA staining showed the tendon was violet, the bone was pink, cartilage was violet, and other tissues were purple. However, the intracellular structures appeared not very well stained. The collagen, elastin and muscular structures in the stained slices were observed under a confocal microscope. Conclusion The commonly used histology staining methods can be used to stain the ultrathin sheet plastination slices. The staining provides a better observation of various tissues in the slice than the unstained slice. After staining, those autofluorescent structures in the plastinated slice are detectable under a confocal microscope.
超薄生物塑化 / 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 / 人
Ultrathin sheet plastination / HE staining / Verhoeff-Van Gieson staining / Methylene blue and azureⅡ staining / Human
[1]Diao Y, Liang L, Yu C, et al. Is there an identifiable intact medial wall of the cavernous sinus? Macro and microscopic anatomical study using sheet plastination[J]. Neurosurgery, 2013, 73(1): 106-109.
[2]Liu M, Chen S, Liang L et al. Microcomputed tomography visualization of the cricoarytenoid joint cavity in cadavers[J]. J Voice, 2013,27(6):778-785.
[3]Zhang ShX, Liu Zh J, He GCh. 3D thin sectional anatomy of sella turcica region with plastination[J]. Acta anatomica sinica, 1998,29(4):337-341.(in Chinese)
张绍祥, 刘正津, 何光篪. 蝶鞍区塑化薄片断层解剖学研究[J]. 解剖学报, 1998, 29(4):337-341.
[4]Zhang BY, Liang L, Liu M, et al. Comparison between the Biodur and PlastMat epoxy mixture[J]. Chinese Journal of Anatomy, 2013, 36(5):948-951. (in Chinese)
张宝营,梁亮,刘明,等.薄层塑化技术要点和进口与国产环氧生物塑化材料的比较[J].解剖学杂志,2013, 36(5):948-951.
[5]Sora MC, Cook P. Epoxy plastination of biological tissue: E12 technique[J]. J Int Soc Plastination, 2007, 22(1): 31-39.
[6]Sora MC. Epoxy plastination of biological tissue: E12 ultra-thin technique[J]. J Int Soc Plastination, 2007, 22(1): 40-45.
[7]Helga F. Staining of different tissues in thick epoxy resin-impregnated sections of human fetuses[J].Stain Technology,1989,64(2): 75-79.
[8]Phillips M, Nash L, Barnett R, et al. The use of confocal microscopy for the examination of E12 sheet plastinated human tissue[J]. J Int Soc Plastination, 2002,17(1): 12-16.
[9]Nash L, Nicholson H, Lee AS, et al. Configuration of the connective tissue in the posterior atlanto-occipital interspace: a sheet plastination and confocal microscopy study[J]. Spine, 2005, 30(12): 1359-1366.
颅中窝外侧区微创外科的应用解剖学研究
/
〈 |
|
〉 |