一种用于溶酶体中酸性磷酸酶活性的X射线微分析的改进方法。
An improved procedure for the X-ray microanalysis of acid phosphatase activity in lysosomes.
作者信息
van Dort J B, Zeelen J P, de Bruijn W C
出版信息
Histochemistry. 1987;87(1):71-7. doi: 10.1007/BF00518727.
The useful detection of acid phosphatase activity with cerium as a capturing agent is confirmed. By introducing a freeze step in combination with a preincubation, reliably localized, lysosomal precipitates are obtained and aspecific ones prevented. Short (t less than 1 h) postfixation with either OsO4 plus K4Fe (CN)6 or OsO4 plus aminotriazole, added to lysosomal cerium localization a high membrane contrast. The detection of cerium by X-ray microanalysis is improved by a better spectral separation of the osmium (M alpha) and cerium (L alpha) peaks.
证实了以铈作为捕获剂对酸性磷酸酶活性进行有效检测。通过引入冷冻步骤并结合预温育，可获得定位可靠的溶酶体沉淀物，并防止非特异性沉淀物的产生。用四氧化锇加亚铁氰化钾（K4Fe(CN)6）或四氧化锇加氨基三唑进行短时间（t小于1小时）的后固定，可增加溶酶体铈定位时的膜对比度。通过更好地分离锇（Mα）和铈（Lα）峰的光谱，改进了通过X射线微分析对铈的检测。
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