解剖学报

解剖学报 ›› 2014, Vol. 45 ›› Issue (3): 416-420.doi: 10.3969/j.issn.0529-1356.2014.03.022

• 技术方法 • 上一篇    下一篇

组织化学染色技术在超薄生物塑化标本中的应用

张宝营1 梁亮1 乔明亮1 张梦楠1 武艳1* 张铭2*   

  1. 1.首都医科大学解剖学教研室，北京 100069; 2. 新西兰奥塔哥大学解剖学系，达尼丁 9016
  • 收稿日期:2013-11-20 修回日期:2014-01-16 出版日期:2014-06-06 发布日期:2014-06-06
  • 通讯作者: 武艳 张铭 E-mail:yanwu@ccmu.edu.cn
  • 基金资助:

    颅中窝外侧区微创外科的应用解剖学研究

Histochemical staining techniques applied in ultrathin sheet plastination slice

ZHANG Bao-ying1 LIANG Liang1 QIAO Ming-liang1 ZHANG Meng-nan1 WU Yan 1* ZHANG Ming 2*   

  1. 1.Department of Anatomy, Capital Medical University, Beijing 100069,China;2. Department of Anatomy，University of Otago, Dunedin 9016
  • Received:2013-11-20 Revised:2014-01-16 Online:2014-06-06 Published:2014-06-06
  • Contact: WU Yan ZHANG Ming E-mail:yanwu@ccmu.edu.cn
  • Supported by:

    Minimally-invasive surgical anatomy of the lateral part of the middle cranial fossa

摘要：

 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。

关键词: 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

Abstract:

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.

Key words: Ultrathin sheet plastination, HE staining, Verhoeff-Van Gieson staining, Methylene blue and azureⅡ staining, Human