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Abstract
Recent work on biological electron energy-loss spectroscopy (EELS) in the analytical electron microscope has focused on measuring concentrations of the important elements, calcium and phosphorus. Analysis of the light elements, for which EELS was originally thought to be best suited, has so far not yielded much useful information from biological specimens. Whereas low-loss fine structure has been analyzed successfully to measure distributions of water and protein in frozen-hydrated cells, core-edge fine structure cannot be recorded at sufficiently low dose to obtain useful chemical information. However, the atomic ratios of the light elements (carbon, nitrogen and oxygen) obtained from EELS are less dependent on electron dose. Together with sulfur (in proteins) and phosphorus (in nucleic acids, phospholipids and phosphates) these ratios can help determine the types and proportions of compounds that are present at the subcellular level. Here we re-examine the potential for making these kinds of measurements.
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