EELS quantification near the single-atom detection level

Krivanek, Ondrej L.; Mory, C.; Tencé, Marcel; Colliex, C.

doi:10.1051/mmm:0199100202-3025700

Cited by 70 publications

(24 citation statements)

References 9 publications

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“…STEM͞EELS has recently gained attention as a near-atomic-resolution structural and nanoanalytical tool for studying materials (14)(15)(16), but its application to biological molecules has been largely precluded by the difficulty of extracting information from very weak signals. This study demonstrates that high-efficiency EELS can detect as few as 10 atoms of phosphorus bound to an individual protein assembly.…”

mentioning

confidence: 99%

Phosphorylation and subunit organization of axonal neurofilaments determined by scanning transmission electron microscopy

Leapman

Gallant

Reese

et al. 1997

Proc. Natl. Acad. Sci. U.S.A.

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Phosphorylation plays a critical role in controlling the function of cytoskeletal assemblies but no direct method yet exists to measure the phosphorylation state of proteins at the level of individual molecules and assemblies. Herein, we apply scanning transmission electron microscopy in combination with electron energy loss spectroscopy to measure the distributions of mass and phosphorus in neurofilaments (NFs) isolated from the squid giant axon. We find that native squid NFs, in contrast to typical reconstituted intermediate filaments, are a relatively homogeneous population containing only eight coiled-coil dimers per cross section. The measured stoichiometry of ϳ1:1 for light͞heavy peptides strongly suggests that squid NFs are composed of heterodimers. Furthermore, each heavy chain of the dimers carries at least 100 phosphate groups and is, therefore, near-maximally phosphorylated. These results also demonstrate that scanning transmission electron microscopy combined with electron energy loss spectroscopy at the nanometer scale is capable of characterizing the level and distribution of phosphorylation in individual mass-mapped protein assemblies.

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mentioning

confidence: 99%

Phosphorylation and subunit organization of axonal neurofilaments determined by scanning transmission electron microscopy

Leapman

Gallant

Reese

et al. 1997

Proc. Natl. Acad. Sci. U.S.A.

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“…For these large n values the optimal aperture angle decreases, thus balancing the effect of the chromatic aberration and the diffraction disc. [33,34]. For both instruments the ultimate limit is imposed by signal/noise considerations [35].…”

Section: Physics Abstractsmentioning

confidence: 99%

Elemental mapping using an imaging energy filter: image formation and resolution limits

Berger

Kohl

1992

Microsc. Microanal. Microstruct.

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“…An improvement to this approach would require cross correlation of the spectra and multiple least squares fitting. 3. Results.…”

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confidence: 99%

Spatially resolved electron energy-loss near-edge structure analysis of a near Σ = 11 tilt boundary in sapphire

Bruley

1993

Microsc. Microanal. Microstruct.

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Abstract. 2014 Spatially resolved electron energy-loss spectra have been recorded from the same E = 11 grain boundary of a high purity sapphire (03B1 -Al2O3) bi-crystal that had previously been examined by high resolution electron microscopy. The energy-loss near-edge structure (ELNES) on the aluminium L2,3 edge of the bulk 03B1 -Al2O3 is consistent with previously published X-ray absorption near-edge spectra (XANES). The presence of the grain-boundary introduces additional fine structure on the L2,3 edge, which is extracted from the dominant bulk spectrum using the chemical-standard "difference" method. A comparison with various spectral "fingerprints" suggests that the structural environment of the Al cations within the boundary plane has a reduced point-group symmetry and coordination number. Quantitative analyses of spectral intensities indicate that the equivalent of a full monolayer of Al cations is involved in the structural change at the boundary. These experimental observations are consistent with a recently calculated low energy relaxed grain-boundary structure.

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EELS quantification near the single-atom detection level

Cited by 70 publications

References 9 publications

Phosphorylation and subunit organization of axonal neurofilaments determined by scanning transmission electron microscopy

Phosphorylation and subunit organization of axonal neurofilaments determined by scanning transmission electron microscopy

Elemental mapping using an imaging energy filter: image formation and resolution limits

Spatially resolved electron energy-loss near-edge structure analysis of a near Σ = 11 tilt boundary in sapphire

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