Biological electron energy loss spectroscopy in the field-emission scanning transmission electron microscope.
The dedicated scanning transmission electron microscope (STEM) combined with parallel electron energy loss spectroscopy (EELS) provides a very sensitive means of detecting specific elements in small structures. EELS is more sensitive than optimized energy-dispersive X-ray spectroscopy by a factor of about three for calcium. Measurement of such low concentrations requires special processing methods such as difference-acquisition techniques and multiple least squares procedures for fitting reference spectra. By analyzing data recorded at each pixel in a spectrum-image it is possible to map quantitatively the elemental distributions in a specimen. It is possible to prepare cryosections that are sufficiently thin to avoid excessive plural inelastic scattering so analysis can be performed at 100 keV beam energy. Under optimal conditions, a resolution of 10 nm and detection limits of a few atoms are achievable for elements such as calcium, phosphorus and iron. In the field emission STEM certain types of chemical information can be extracted from biological specimens. Valence EELS has been exploited to measure water distributions in frozen hydrated cryosections.
Leapman
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Biomedical Engineering & Instrumentation Program, NCRR, National Institutes of Health, Bethesda, MD 20892, USA.
Sun
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Hunt
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Andrews
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D000818:Animals; D002118:Calcium / Q000032:analysis*; D002525:Cerebellar Cortex / Q000648:ultrastructure*; D003712:Dendrites / Q000648:ultrastructure; D004721:Endoplasmic Reticulum / Q000648:ultrastructure; D005615:Freezing; D008433:Mathematics; D051379:Mice; D017348:Microscopy, Electron, Scanning Transmission / Q000295:instrumentation / Q000379:methods*; D008928:Mitochondria / Q000648:ultrastructure; D009474:Neurons / Q000648:ultrastructure*; D011689:Purkinje Cells / Q000648:ultrastructure*; D012680:Sensitivity and Specificity; D013057:Spectrum Analysis
D002118:Calcium