Porcine Intestinal Mast Cells. Evaluation of Different Fixatives for Histochemical Staining Techniques Considering Tissue Shrinkage
Porcine Intestinal Mast Cells. Evaluation of Different Fixatives for Histochemical Staining Techniques Considering Tissue Shrinkage
复制标题
猪肠肥大细胞。
DOI：
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发表时间：
2013
影响因子：
2
通讯作者：
Johanna Plendl
中科院分区：
文献类型：
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作者：
Juliane Rieger;Sven Twardziok;H. Huenigen;R. Hirschberg;Johanna Plendl
肥大细胞（包括猪肥大细胞）的染色主要依赖于固定和染色技术。在猪中，粘膜和粘膜下MC不染色或在福尔马林固定后仅轻微染色。一些固定方法特别推荐用于MC染色，例如用Carnoy或铅盐固定。锌盐固定法（Zinc salt fixation，简称ZIF）是一种保存固定敏感抗原的有效方法。本研究的目的是建立一种可靠的组织学方法来计数猪睾丸中的MC。为此，在猪粘膜以及小肠和大肠粘膜下层中评价了不同的组织固定和染色方法，这些方法也允许潜在的后续免疫组织化学研究。将组织固定在Carnoy、醋酸铅、硝酸铅、Zamboni和NaF中，随后用多色亚甲蓝、阿辛蓝或甲苯胺蓝染色。我们的研究第一次揭示了，BMPF，一种重金属固定剂，保留了猪MC的异染性染色。Zamboni固定法不适用于猪小肠MC的组织化学观察。所有其他受试固定剂均适用。阿辛蓝和甲苯胺蓝共染小肠杯状细胞，这使得初步鉴定MC变得困难。事实证明，多色亚甲基蓝是最佳染色法。为了比较不同固定方法的MC计数结果，考虑了组织收缩。由于即使相同的固定也会导致小肠和大肠组织之间的收缩差异，因此必须计算每个单一固定和肠定位的不同因子。组织收缩率在19%~ 57%之间变化，大肠组织收缩率以BMF固定后最高，小肠组织收缩率以醋酸铅固定后最低。我们的研究强调，MC计数结果的数据使用不同的固定技术只能进行比较，如果各自的研究immunohistochemistry收缩因子已被确定和量化结果进行了相应的调整。
Staining of mast cells (MCs), including porcine ones, is critically dependent upon the fixation and staining technique. In the pig, mucosal and submucosal MCs do not stain or stain only faintly after formalin fixation. Some fixation methods are particularly recommended for MC staining, for example the fixation with Carnoy or lead salts. Zinc salt fixation (ZSF) has been reported to work excellently for the preservation of fixation-sensitive antigens. The aim of this study was to establish a reliable histological method for counting of MCs in the porcine intestinum. For this purpose, different tissue fixation and staining methods that also allow potential subsequent immunohistochemical investigations were evaluated in the porcine mucosa, as well as submucosa of small and large intestine. Tissues were fixed in Carnoy, lead acetate, lead nitrate, Zamboni and ZSF and stained subsequently with either polychromatic methylene blue, alcian blue or toluidine blue. For the first time our study reveals that ZSF, a heavy metal fixative, preserves metachromatic staining of porcine MCs. Zamboni fixation was not suitable for histochemical visualization of MCs in the pig intestine. All other tested fixatives were suitable. Alcian blue and toluidine blue co-stained intestinal goblet cells which made a prima facie identification of MCs difficult. The polychromatic methylene blue proved to be the optimal staining. In order to compare MC counting results of the different fixation methods, tissue shrinkage was taken into account. As even the same fixation caused shrinkagedifferences between tissue from small and large intestine, different factors for each single fixation and intestinal localization had to be calculated. Tissue shrinkage varied between 19% and 57%, the highest tissue shrinkage was found after fixation with ZSF in the large intestine, the lowest one in the small intestine after lead acetate fixation. Our study emphasizes that MC counting results from data using different fixation techniques can only be compared if the respective studyimmanent shrinkage factor has been determined and quantification results are adjusted accordingly.
影响因子：
14.2
作者：
Stone, Kelly D.;Prussin, Calman;Metcalfe, Dean D.
通讯作者：
Metcalfe, Dean D.
DOI：
10.1152/ajpgi.00081.2009
发表时间：
2010-03-01
影响因子：
4.5
作者：
Smith, Feli;Clark, Jessica E.;Moeser, Adam J.
通讯作者：
Moeser, Adam J.