A novel method for the visualization of the in situ mucus layer in rat and man
A novel method for the visualization of the in situ mucus layer in rat and man
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DOI：
10.1042/cs19980081
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发表时间：
1998-07-01
期刊：
影响因子：
6
通讯作者：
Allen, A
中科院分区：
文献类型：
--
作者：
Jordan, N;Newton, J;Allen, A
观察到的胃粘液屏障厚度差异很大，甚至看起来不连续，这取决于所使用的方法。在这里，我们描述了一种改进的高碘酸希夫/阿尔辛蓝染色技术的发展和应用，用于胃粘膜冰冻切片。这项技术首次能够保存和显示粘连的胃粘液层和下粘膜的全层厚度。2.在设计这一新的方法时，我们选择了对粘液层影响最小的程序。采用的方法是快速冷冻、全胃冷冻切片，然后进行短暂的乙醇预处理(100%乙醇中10min)，高碘酸希夫/阿尔西安蓝延长染色时间(分别为15min和2.5h)，温和的后处理(37℃多聚甲醛蒸气45min)，以及使用水溶性贴剂。在大鼠胃粘膜(高碘酸席夫蓝染色)和人胃窦活检组织(高碘酸席夫蓝染色)表面观察到一层连续的粘液层。大鼠的平均数(S.D.)沿胃窦至食管轴的粘液厚度(组织学上分为四个区域，从A到D)为：A，166(47)微米；B，179(48)微米；C，184(50)微米；D(胃顶部的非腺复层上皮)，无。人胃窦活检的平均值(S.D.)粘液厚度为144(52)微米4，这项新技术使可视化和精确测量大鼠和人的胃粘液层厚度成为可能。粘附层在大鼠腺胃和人胃窦内连续存在。在大鼠的验证性实验中，粘液厚度的平均测量值是传统组织学方法的两倍，粘液层呈现不连续和斑片状。然而，它们在未固定组织和大鼠体内制剂中看到的厚度值范围内。
1, The observed thickness of the gastric mucus barrier varies widely, even appearing discontinuous, depending on the methods used. Here we describe the development and application of a modified periodic acid Schiff/Alcian Blue staining technique for use on cryostat sections of gastric mucosa. This technique for the first time enables the preservation and visualization of the full thickness of the adherent gastric mucus layer and the underlying mucosa.2, In designing this novel method we have selected those procedures which would result in the least alteration to the mucus layer. The methods used were snap freezing, cryostat sectioning of the whole stomach followed by brief ethanol pretreatment (10 min in 100% ethanol), a prolonged staining with periodic acid Schiff/Alcian Blue (15 min and 2.5 h respectively), a gentle post-fixation (45 min paraformaldehyde vapour at 37 degrees C) and the use of a water-soluble mountant.3. A continuous, adherent mucus layer was observed over the surface of the rat gastric mucosa (periodic acid Schiff/Alcian Blue stained) and human gastric antral biopsies (periodic acid Schiff stained). In the rat the mean (S.D.) mucus thickness measurements along the antrum to oesophageal axis (which; was divided histologically into four regions, A to D) were: A, 166 (47) mu m; B, 179 (48) mu m; C, 184 (50) mu m; D (the non-glandular stratified epithelium at the top of the stomach), Absent. In human gastric antral biopsies the mean (S.D.) mucus thickness was 144 (52) mu m.4, This new technique has enabled the visualization and precise measurement of thickness of the gastric mucus layer in rat and man. The adherent gastric mucus layer was observed to be continuous in the rat glandular stomach and human antrum. In validation experiments in rat the mean mucus thickness measurements were found to be twice those measured by conventional histological techniques, in which the mucus layer appeared discontinuous and patchy. However, they were within the range of thickness values seen in unfixed tissues and in the rat in vivo preparation.