Image‐EELS: Simultaneous recording of multiple electron energy‐loss spectra from series of electron spectroscopic images

Körtje, Karl‐Heinz

doi:10.1111/j.1365-2818.1994.tb03463.x

Cited by 31 publications

(20 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Titanium was not detectable in the nucleus of the same cells using SEELS analysis. 10 [3] is one way around this constraint. Our approach sacrifices energy resolution for an increase in the size of the analysed area.…”

Section: Analysis Of Nafionmentioning

confidence: 99%

“…The technique, known as Spectrum Imaging [1], Image Spectrum [2] or Image-EELS [3], records energy specific elemental loses as a grey scale within an image array from one or more elements within a large area for several images over a defined electron energy loss range. In addition to elemental information, the digital image sequence also contains grey level intensity variations corresponding to information about local changes in mass/thickness and the non-specific background energy contributions, which can be determined by modelling the local energy spectrum.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Electron Spectroscopic Imaging of Organic Compounds Using PC-Based Energy Sequence Imaging Software

Johnson

Mairs

Gaze

et al. 1995

Microsc. Microanal. Microstruct.

View full text Add to dashboard Cite

Abstract. 2014 We have designed, characterised and tested Windows based software packages for rapid elemental microanalysis using a sequence of energy filtered transmission electron microscopic images which combines electron spectroscopic imaging with electron energy loss spectroscopy (Image-EELS). Image sequences containing up to 45 images of 512 x 512 pixels were recorded rapidly over an appropriate range of the energy spectrum for an analysis of boron, nitrogen, titanium and fluorine. Energy loss information was extracted from local variations in the energy loss spectrum with good energy resolution (5 eV) and a spatial resolution of 2 nm for the determination of the relative concentration of elements within a region of interest by grey level integration in consecutive images of an energy loss sequence, using the least mean squares method to calculate the parameters for an A · e-r background model. The methods were used to define the optimum collections conditions and practical limitations of the system for qualitative and semi-quantitative elemental analysis relevant to biological specimens. Qualitative elemental analysis of images from Lowicryl HM23-embedded ultra-thin section of a boric acid polymer showed that boron was not distributed evenly. Semi-quantitative elemental analysis of the fluorine-containing organic polymer Nafion suggested a linear relationship between fluorine energy loss intensity, image area and specimen thickness. These techniques were applied to an examination of the reaction between purified RNA and the novel titanium-containing anti-cancer agent MKT-4, showing that ratios calculated from the cumulative electron energy loss intensity increase within regions of interest as a function of grey level intensity for titanium and nitrogen are directly proportional to the incubation concentration of the drug with RNA. Expression of data in this way is shown to be a useful indicator of the relative drug distribution within specific intracellular compartments of human cancer cells incubated with MKT-4 in vitro.

show abstract

Section: Analysis Of Nafionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Electron Spectroscopic Imaging of Organic Compounds Using PC-Based Energy Sequence Imaging Software

Johnson

Mairs

Gaze

et al. 1995

Microsc. Microanal. Microstruct.

View full text Add to dashboard Cite

show abstract

“…For this, inelastically scattered electrons with a defined energy-loss obtained by interaction with the electrons of the specimen can be analyzed. These electrons are selected with a spectrometer and can be used for electron spectroscopic imaging (ESI), for electron energy-loss spectroscopy (EELS) (Reimer et al, 19921, and for Image-EELS (Kortje, 1994). The photographs were taken in structure-sensitive contrast mode which shows a reversed contrast (Reimer et al, 1992)…”

Section: Embedding Proceduresmentioning

confidence: 99%

Comparative methodological investigations on the cytochemical localization of calcium in brain and inner ear of cichlid fish

Vöhringer

Nindl

Aich

et al. 1995

Microscopy Res & Technique

View full text Add to dashboard Cite

Four different methods for calcium precipitation are compared in the optic tectum and the inner ear of the cichid fish, Oreochromis mossambicus. Several parameters are investigated concerning their influences on the reaction product. Three procedures (bichromate, fluoride, and oxalate-pyroantimonate) produce fine-grained deposits, often flocculent in the latter method. The fourth method (potassium-pyroantimonate) generates predominantly coarse-grained reaction product. The calcium content of the deposits is always proven with energy-filtering transmission electron microscopy (EFTEM). In both tissues fine-grained reaction product is found in endoplasmic reticulum and synaptic vesicles, and in addition in some mitochondria and at the cytoskeleton. The coarse-grained deposits of the potassium-pyroantimonate method have a more unspecific distribution. This is the only method which produces extracellular deposits in the inner ear, whereas in the optic tectum extracellular precipitates are always present except with the oxalate-pyroantimonate procedure. Two factors have an influence on the reaction product: the duration of fixation and the type of resin. The prolongation of the fixation time up to 24 hours leads to an increase of the reaction product, which also becomes coarse-grained. These observations are corroborated by quantification with image analysis. Furthermore the use of an epoxy resin compared to acrylic resins decreases the amount of reaction product produced. We show that the application of several methods is meaningful in order to understand the calcium properties of the investigated tissue, but it is necessary to optimize a certain method for a given tissue.

show abstract

“…While the 'Imaging-spectrum' and 'Image-EELS' methods (Lavergne et al, 1992(Lavergne et al, , 1994Körtje, 1994) yield spectra which can be processed further, videodensitometry yields a particular sort of plot which gives only qualitative information. However, videodensitometry is presently the only method which allows the detection of small local concentrations of biologically relevant elements directly from micrographs where the instrumentation for Image-EELS is not affordable.…”

Section: I D E O D E N S I T O M E T Ry F O R M I C Roa Na Lys I Smentioning

confidence: 99%

“…Some peculiarities of this method will be discussed here further to verify the reliability of the results. Alternative methods of densitometric image evaluations have been published, called 'Imaging-Spectrum' (Lavergne et al, 1992;Lavergne et al, 1994) and 'Image-EELS' (Körtje, 1994), respectively, to distiguish them from the 'Spectrum-Imaging' method (Hunt & Williams, 1991) in which images are created by pixel-to-pixel evaluation of parallel EEL spectra sampled in the STEM.…”

Section: Introductionmentioning

confidence: 99%

Videodensitometric analysis of electron spectroscopic micrographs — a tool for detection of biologically relevant elements with high resolution

Door¹,

Breitig

Martin³

1997

Journal of Microscopy

View full text Add to dashboard Cite

SummaryElectron energy-loss spectroscopic imaging (ESI) yields highresolution, element-sensitive images. However, ESI suffers from difficulties in distinguishing element-specific and background contributions. New methods have therefore been introduced which use grey-level measurements in micrographic images for a more accurate detection of element distributions. A videodensitometric method allowed the detection of low phosphorus levels in axoplasmic neurofilaments of squid giant axons. Here we further verify these results by investigating the relationship of videodensitometry and electron energy-loss spectroscopy (EELS), particularly considering the peculiarities of these methods in terms of automatic background correction and representation of the results. Six biological specimens and two nonbiological specimens were examined both by EELS and by videodensitometry. In all cases comparable results were obtained. The overlapping P L2,3 and S L2,3 ionization edges could clearly be recognized individually by both methods, and controls showed that mass density variations within the specimens did not impair elemental analysis. Additional evidence supporting the detection of phosphorylation sites in squid neurofilaments was obtained in both EELS and videodensitometric measurements of neurofilament-enriched pellets and of aggregated axoplasmic particles. Thus, videodensitometry appears to be a useful tool for an improved exploration of the full imaging capabilities of energy filtering electron microscopy.

show abstract

Image‐EELS: Simultaneous recording of multiple electron energy‐loss spectra from series of electron spectroscopic images

Cited by 31 publications

References 15 publications

Electron Spectroscopic Imaging of Organic Compounds Using PC-Based Energy Sequence Imaging Software

Electron Spectroscopic Imaging of Organic Compounds Using PC-Based Energy Sequence Imaging Software

Comparative methodological investigations on the cytochemical localization of calcium in brain and inner ear of cichlid fish

Videodensitometric analysis of electron spectroscopic micrographs — a tool for detection of biologically relevant elements with high resolution

Contact Info

Product

Resources

About