Quantitative X-ray Images Calculated on the Basis of Mass and Volume

Quantitative X-ray microanalysis is used to obtain the elemental compositions of tissues and cells. Concentrations are calculated on the basis of volume (mmolL−1 of packed cells) and mass (mmolkg−1 dry weight). Elemental maps are obtained by using a computer to control the position of a beam in an electron microscope and to record the signals from the scanning transmission electron microscope (STEM) and energy dispersive X-ray spectrometer (EDS) X-ray detectors. These X-ray images provide better visualization of elemental distributions than ``spot'' mode analysis by analyzing thousands of spots sequentially. When concentrations are determined on the basis of volume, it is assumed that the section thickness created during cryosectioning, and shrinkage during freeze-drying of a frozen section, are uniform. These assumptions have been examined with a nucleated red blood cell model. Even distributions of cytoplasmic Fe and K, which we observed, can only occur if cryomicrotomy produces sections with smooth surfaces and uniform thickness. In addition, by using bone marrow cryosections we have found that the relative shrinkage between nucleated and non-nucleated cells is similar. Therefore, the assumptions made about volume calculations do appear to be reasonable under the conditions used in this study.