Alterations of mouse lung tissue dimensions during processing for morphometry: A comparison of methods
Alterations of mouse lung tissue dimensions during processing for morphometry: A comparison of methods
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DOI：
10.1152/ajplung.00329.2013
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发表时间：
2014-02-01
影响因子：
4.9
通讯作者：
Ochs, Matthias
中科院分区：
文献类型：
--
作者：
Schneider, Jan Philipp;Ochs, Matthias
保存原始组织尺寸是形态计量学研究的必要先决条件。在组织组织学处理过程中发生的收缩可能严重影响在显微镜和立体学计算下看到的结构外观。因此，必须避免收缩，以便通过应用无偏立体学获得的估计确实是无偏的。本研究探讨了小鼠肺标本在组织学处理过程中组织尺寸的变化。考虑了不同的固定剂和嵌入协议。用4%福尔马林或1.5%戊二醛/1.5%甲醛的混合物固定小鼠肺。组织块按体视学原理取样，包埋石蜡、未经四氧化锇和醋酸铀酰处理的甲基丙烯酸乙二醇酯和脱水前经四氧化锇和醋酸铀酰处理的甲基丙烯酸乙二醇酯。通过对组织切割面尺寸变化的立体测量来研究收缩。结果表明，石蜡包埋时，每肺切口面积收缩约40%，“简单”甲基丙烯酸乙二醇酯包埋时收缩30%
Preservation of original tissue dimensions is an essential prerequisite for morphometric studies. Shrinkage occurring during tissue processing for histology may severely influence the appearance of structures seen under the microscope and stereological calculations. Therefore, shrinkage has to be avoided so that estimates obtained by application of unbiased stereology are indeed unbiased. The present study investigates the alterations of tissue dimensions of mouse lung samples during processing for histology. Different fixatives as well as embedding protocols are considered. Mouse lungs were fixed by instillation of either 4% formalin or a mixture of 1.5% glutaraldehyde/1.5% formaldehyde. Tissue blocks were sampled according to principles of stereology for embedding in paraffin, glycol methacrylate without treatment with osmium tetroxide and uranyl acetate, and glycol methacrylate including treatment with osmium tetroxide and uranyl acetate before dehydration. Shrinkage was investigated by stereological measurements of dimensional changes of tissue cut faces. Results show a shrinkage of the cut face areas of roughly 40% per lung during paraffin embedding, 30% during "simple" glycol methacrylate embedding, and