Improved Swiss-rolling Technique for Intestinal Tissue Preparation for Immunohistochemical and Immunofluorescent Analyses
Improved Swiss-rolling Technique for Intestinal Tissue Preparation for Immunohistochemical and Immunofluorescent Analyses
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DOI：
10.3791/54161
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发表时间：
2016-07-01
影响因子：
1.2
通讯作者：
Yang, Vincent W.
中科院分区：
文献类型：
--
作者：
Bialkowska, Agnieszka B.;Ghaleb, Amr M.;Yang, Vincent W.
了解调节肠上皮细胞动态平衡以及对损伤和再生反应的因素的作用是重要的。目前的文献描述了几种不同的方法来获得用于数据验证的肠组织图像。在这篇文章中，我们描述了一个与小鼠肠道组织的派生和处理有关的通用协议。讨论了肠道组织的适当固定和增强肠上皮形态的Swiss-roll技术。在获得切片后显示完整肠道结构特征的石蜡包埋块时，对包埋的肠道组织的定位、处理和重定向至关重要。瑞士卷曲技术有助于对所检查的完整肠道或结肠切片进行组织学评估。在整个长度上定量测量肠道炎症和肿瘤发生时，区分肠道结构特征的能力可能是至关重要的。最后，石蜡包埋切片是使用免疫组织化学和免疫荧光检测方法进行稳健处理的理想选择。非荧光免疫组织化学切片提供了组织的生动图像，详细描述了不同的细胞结构特征，但不能为细胞内共定位实验提供灵活性。多个荧光通道可以适当地与免疫荧光检测一起用于共定位实验，为机制研究提供支持。
Understanding the role of factors that regulate intestinal epithelial homeostasis and response to injury and regeneration is important. The current literature describes several different methodological approaches to obtain images of intestinal tissues for data validation. In this paper, we delineate a common protocol relating to the derivation and processing of mouse intestinal tissues. Proper fixation of intestinal tissues and Swiss-roll techniques that enhance intestinal epithelial morphology are discussed. Postresection processing and reorientation of embedded intestinal tissues are critical in obtaining paraffin-embedded blocks that display intact intestinal structural features after sectioning. The Swiss-rolling technique helps in histological assessment of the complete intestinal or colonic sections examined. An ability to differentiate intestinal structural features can be vital in quantitative measurements of intestinal inflammation and tumorigenesis along the entire length. Finally, paraffin-embedded sections are ideal for robust processing using both immunohistochemical and immunofluorescent detection methods. Nonfluorescent immunohistochemical sections provide a vibrant image of the tissue detailing different cellular structural features but do not provide flexibility for intracellular co-localization experiments. Multiple fluorescent channels can be appropriately utilized with immunofluorescent detection for co-localization experiments, lending support to mechanistic studies.