Horobin R W, Gerrits P O, Wright D J
Department of Biomedical Science, University, Sheffield, U.K.
J Microsc. 1992 May;166(Pt 2):199-205. doi: 10.1111/j.1365-2818.1992.tb01518.x.
Glycol methacrylate (GMA) sections of animal tissues were stained with a group of twenty-seven reagents of very varied chemical characteristics. The artefactual background staining of the resin was found to be dependent on the hydrophilic/lipophilic character of the staining reagent, as estimated from the logarithm of its octanol-water partition coefficient (log P). Intense background staining occurred with lipophilic stains, whose log P greater than 2. In keeping with this, use of GMA semi-permeable membranes for enzyme histochemistry failed to give staining when using a lipophilic substrate, probably because the substrate was trapped in the membrane. An analysis of other routine histochemical stains--in terms of the probable occurrence of high resin background staining and low tissue sensitivity--is made. A numerical guide is provided to help avoid artefacts resulting from hydrophobic and size effects. Note: small, hydrophilic reagents (log P less than 0; molecular weight less than 550 Da) are least likely to show either type of artefact. Conversely, reagents which are lipophilic, or/and of intermediate size (log P greater than 2; 550 less than ionic weight less than 1000 Da), give strong background staining.
动物组织的甲基丙烯酸乙二醇酯（GMA）切片用一组具有非常不同化学特性的27种试剂进行染色。发现树脂的人为背景染色取决于染色试剂的亲水/亲脂特性，这是根据其辛醇 - 水分配系数的对数（log P）估算的。亲脂性染色剂会出现强烈的背景染色，其log P大于2。与此一致的是，在酶组织化学中使用GMA半透膜时，使用亲脂性底物未能产生染色，可能是因为底物被困在膜中。对其他常规组织化学染色剂进行了分析，考虑了高树脂背景染色和低组织敏感性可能出现的情况。提供了一个数字指南，以帮助避免由疏水和尺寸效应导致的假象。注意：小的亲水性试剂（log P小于0；分子量小于550 Da）最不可能出现任何一种假象。相反，亲脂性的和/或中等大小的试剂（log P大于2；550小于离子重量小于1000 Da）会产生强烈的背景染色。
