Ultrathin sectioning for electron microscopy: the distilled water in the knife trough may extract phosphatase reaction products from the sections.
During an electron microscopical study of the localization of the nucleoside diphosphatase IDPase in Reissner's membrane of the inner ear, it was discovered that the distilled water in the knife trough produced an annoying artefact. It dissolved all the lead phosphate reaction product from the sections, and thus converted a positive phosphatase reactivity to a false negative one. The water in the knife trough had a pH of approximately 5.4. Calculations showed that this is an expected acidity, if CO2 in the air equilibrates with distilled water, and that there is 200,000 times more acid in the trough than necessary to dissolve all the reaction product from a ribbon of ultrathin sections. Experiments showed that the artefact could be avoided by adjusting the pH to neutrality with dilute ammonia.
Rostgaard
J
J
Institute of Medical Anatomy, Department B, University of Copenhagen, Panum Institute, Denmark.
Qvortrup
K
K
D017766:Acid Anhydride Hydrolases; D000818:Animals; D007758:Ear, Inner / Q000201:enzymology / Q000648:ultrastructure; D006168:Guinea Pigs; D006863:Hydrogen-Ion Concentration; D008854:Microscopy, Electron / Q000379:methods*; D008867:Microtomy / Q000379:methods*; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D014867:Water
D014867:Water; D010744:Phosphoric Monoester Hydrolases; D017766:Acid Anhydride Hydrolases; C021446:inosine diphosphatase