Quantitative electron energy loss spectroscopy in biology.
The potential for applying electron energy loss spectroscopy (EELS) in biology is assessed. Some recent developments in instrumentation, spectrometer design, parallel detection and elemental mapping are discussed. Quantitation is demonstrated by means of the spectrum from DNA which gives an elemental ratio for N:P close to the expected value. A range of biologically important elements that can be usefully analyzed by EELS is tabulated and some possible applications for each are indicated. Detection limits and the effects of radiation damage are illustrated by spectra from the protein, insulin, and from the fluorinated amino-acid, histidine. Calcium detectability under optimum conditions may be as low as 1 mmol/kg dry weight. The application of EELS to analysis of cryosectioned adrenomedullary (chromaffin) cells is described in order to help determine the composition of the secretory granule. Water content can be determined from the amount of inelastic scattering as measured by the low-loss spectrum. The nitrogen/phosphorus ratio can be measured to provide information about the relative concentrations of ATP, chromogranin, and catecholamines. Quantitative EELS elemental maps are obtained in the STEM mode from chromaffin cells in order to measure the distribution of light elements.
Leapman
R D
RD
Biomedical Engineering and Instrumentation Branch, National Institutes of Health, Bethesda, Maryland 20892.
Ornberg
R L
RL
D001834:Body Water / Q000032:analysis; D002837:Chromaffin Granules / Q000032:analysis; D003594:Cytoplasmic Granules / Q000032:analysis; D004247:DNA / Q000032:analysis; D004583:Electrons; D004602:Elements / Q000032:analysis*; D013057:Spectrum Analysis
D004602:Elements; D004247:DNA