ADVANTAGEOUS USAGE OF EELS TO DETECT CA IN PYROANTIMONATE STAINING

Potassium pyroantimonate staining has been utilized to localize calcium (Ca) in biological specimens at the electron microscopic level. This method, however, has been criticized by many investigators because of its specificity to Ca. X-ray microanalysis (EDX) of that reaction product, pyroantimonate-Ca, was an attempt to confirm Ca in it. There is a technical probrem in distinguishing Ca and antimony (Sb) by EDX because the characteristic X-ray emission energies of Sb and Ca are too close each other to be distinguished.Therefore, the reaction product localized in the cytoplasm and over the plasma membrane of type II cells in taste buds of four wild Japanese monkeys was analysed with an electron energy loss spectroscope (EELS) combined with STEM image mode. Ca peak was recognized in the EELS spectrum with the addition of 2% K-pyroantimonate to 4% glutaraldehyde. Similar addition of K-pyroantimonate to osmium tetroxide was not essential to preserve Ca for EELS survey. Thus, EELS appeared to be more advantageous as compared to EDX in detection of Ca in histochemical reaction product.

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