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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
Pt 2
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pubmed:dateCreated |
1992-5-18
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pubmed:abstractText |
The combination of scanning transmission electron microscopy (STEM) and parallel-detection energy-loss spectroscopy (EELS) was used to detect specific bound elements within macromolecules and macromolecular assemblies prepared by direct freezing. After cryotransferring and freeze-drying in situ, samples were re-cooled to liquid nitrogen temperature and low-dose (about 10(3) e/nm2) digital dark-field images were obtained with single-electron sensitivity using a beam energy of approximately 100 keV and a probe current of approximately 5 pA. These maps provided a means of characterizing the molecular weights of the structures at low dose. The probe current was subsequently increased to about 5 nA in order to perform elemental analysis. The 320 copper atoms in a keyhole limpet haemocyanin molecule (mol.wt = 8 MDa) were detected with a sensitivity of +/- 30 atoms in an acquisition time of 200 s. Phosphorus was detected in an approximately 10-nm length of single-stranded RNA contained in a tobacco mosaic virus particle (mol.wt = 130 kDa/nm) with a sensitivity of +/- 25 atoms. Near single-atom sensitivity was achieved for the detection of iron in one haemoglobin molecule (mol.wt = 65 kDa, containing four Fe atoms). Such detection limits are only feasible if special processing methods are employed, as is demonstrated by the use of the second-difference acquisition technique and multiple least-squares fitting of reference spectra. Moreover, an extremely high electron dose (about 10(10) e/nm2) is required resulting in mass loss that may be attributable to 'knock-on' radiation damage.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Ferritins,
http://linkedlifedata.com/resource/pubmed/chemical/Hemocyanin,
http://linkedlifedata.com/resource/pubmed/chemical/Hemoglobins,
http://linkedlifedata.com/resource/pubmed/chemical/Iron,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphorus
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pubmed:status |
MEDLINE
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pubmed:month |
Feb
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pubmed:issn |
0022-2720
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
165
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
225-38
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:1564721-Animals,
pubmed-meshheading:1564721-Ferritins,
pubmed-meshheading:1564721-Hemocyanin,
pubmed-meshheading:1564721-Hemoglobins,
pubmed-meshheading:1564721-Iron,
pubmed-meshheading:1564721-Microscopy, Electron, Scanning,
pubmed-meshheading:1564721-Mollusca,
pubmed-meshheading:1564721-Phosphorus,
pubmed-meshheading:1564721-Spectrum Analysis,
pubmed-meshheading:1564721-Tobacco Mosaic Virus
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pubmed:year |
1992
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pubmed:articleTitle |
Characterization of biological macromolecules by combined mass mapping and electron energy-loss spectroscopy.
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pubmed:affiliation |
Biomedical Engineering and Instrumentation Program, NCRR, National Institutes of Health, Bethesda, MD 20892.
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pubmed:publicationType |
Journal Article
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