Edwards Kimberley, Griffiths David, Morgan John, Pitman Ruth, von Ruhland Christopher
Department of Histopathology, Cardiff and Vale NHS Trust, UK.
Nephrol Dial Transplant. 2009 Feb;24(2):400-3. doi: 10.1093/ndt/gfn504. Epub 2008 Sep 2.
Establishing glomerular basement membrane (GBM) thickness is important in the diagnosis of some renal diseases. It is widely believed that GBM thickness varies according to the processing method, yet there appear to be no published data to support this. In this study we aimed to determine whether the choice of intermediate solvent and embedding resin influenced the thickness of the GBM and to assess the magnitude of any such effect.
Subcapsular renal cortical rat tissue was processed for electron microscopy using four different intermediate solvents [propylene oxide (PO), xylene (Xyl), acetone (Ac) or ethanol (Et)], and three epoxy resins [Araldite (Aral), TAAB embedding resin (TER) or TAAB low viscosity resin (TLV)]. GBM thickness was estimated by the orthogonal intercept method.
Compared with PO-Aral embedding, GBM was significantly thinner in Xyl-Aral, Ac-Aral and Xyl-TER, and significantly thicker in PO-TER-, Ac-TER-, Et-TER- and PO-TLV-embedded tissue (P < 0.05 in all cases). No significant difference was seen with Xyl-TLV- and Ac-TLV-embedded tissue. Et-Aral and Et-TLV embedding resulted in poor quality blocks.
The results suggest that the choice of both intermediate solvent and resin influences shrinkage of the GBM during TEM processing.
确定肾小球基底膜（GBM）厚度对某些肾脏疾病的诊断很重要。人们普遍认为GBM厚度会因处理方法而异，但似乎尚无已发表的数据支持这一点。在本研究中，我们旨在确定中间溶剂和包埋树脂的选择是否会影响GBM的厚度，并评估任何此类影响的程度。
使用四种不同的中间溶剂[环氧丙烷（PO）、二甲苯（Xyl）、丙酮（Ac）或乙醇（Et）]和三种环氧树脂[Araldite（Aral）、TAAB包埋树脂（TER）或TAAB低粘度树脂（TLV）]对大鼠肾皮质被膜下组织进行电子显微镜处理。通过正交截距法估计GBM厚度。
与PO-Aral包埋相比，Xyl-Aral、Ac-Aral和Xyl-TER中的GBM明显更薄，而在PO-TER、Ac-TER、Et-TER和PO-TLV包埋的组织中GBM明显更厚（所有情况均P < 0.05）。Xyl-TLV和Ac-TLV包埋的组织未见显著差异。Et-Aral和Et-TLV包埋导致组织块质量差。
结果表明，中间溶剂和树脂的选择均会影响透射电镜处理过程中GBM的收缩。
