An improved procedure for the X-ray microanalysis of acid phosphatase activity in lysosomes |
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| Authors: | J. B. van Dorf J. Ph. Zeelen W. C. de Bruijn |
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| Affiliation: | (1) Department of Histology and Cytology, Hogeschool West-Brabant, Sektor Laboratoriumonderwijs, Postbus 4953, 4803 EZ Breda, The Netherlands;(2) Laboratory for Experimental Electron Microscopy, Pathological Laboratory I, Erasmus University, P.O. Box 1738, 3000 DR Rotterdam;(3) Analytical Microscopy, Laboratory for Electron Microscopy, Rijnsburgerweg 10, 2333 AA Leiden, The Netherlands |
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| Abstract: | Summary 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<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  ) and cerium (L ) peaks. |
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