Walton J R
Australian Institute for Biomedical Research, Marrickville (Sydney) NSW, Australia.
Biotech Histochem. 2004 Oct-Dec;79(5-6):169-76. doi: 10.1080/10520290400015423.
A sensitive bright field/fluorescent histochemical staining method has been developed that reveals endogenous aluminum in subcellular structures. The method, achievable within 30 min, is based on phloxine B and phosphotungstic acid, with ethanol differentiation. Hematoxylin is used for nuclear and fast green FCF for cytoplasmic counterstaining. To test the method's specificity, we incubated living neuroblastoma cells overnight in culture media containing aluminum, calcium, iron, copper or zinc, or no added metal ions. After fixing the cells and applying the staining method, only cultures exposed to aluminum stained magenta. Applying the method to paraffin embedded tissue sections pretreated with one of two chelating agents that remove aluminum demonstrated less magenta staining in the chelated sections than in adjacent unchelated sections. Immersing sections overnight in solutions containing exogenous aluminum had no observable effect on staining for endogenous aluminum; therefore, it is unlikely that any exogenous aluminum present in histological reagents would alter the method's staining results.
已开发出一种灵敏的明场/荧光组织化学染色方法，可揭示亚细胞结构中的内源性铝。该方法基于焰红B和磷钨酸，用乙醇分化，30分钟内即可完成。苏木精用于细胞核复染，固绿FCF用于细胞质复染。为测试该方法的特异性，我们将活的神经母细胞瘤细胞在含有铝、钙、铁、铜或锌的培养基中或不添加金属离子的培养基中培养过夜。固定细胞并应用染色方法后，只有暴露于铝的培养物染成品红色。将该方法应用于用两种去除铝的螯合剂之一预处理的石蜡包埋组织切片，螯合切片中的品红色染色比相邻的未螯合切片少。将切片在含有外源性铝的溶液中浸泡过夜，对内源性铝的染色没有可观察到的影响；因此，组织学试剂中存在的任何外源性铝不太可能改变该方法的染色结果。
