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Biomedical subjects

A L Beckers

Publications and source records attributed to A L Beckers.

6 recordsLinked to original sources

New developments and applications in quantitative electron spectroscopic imaging of iron in human liver biopsies.

Reliable iron concentration data can be obtained by quantitative analyses of image sequences, acquired by electron spectroscopic imaging. A number of requirements are formulated for the successful application of this recently developed in situ quantitative type of analysis. A demonstration of the procedures is given. By application of the technique it is established that there are no significant differences in the average iron loading of structures analysed in liver parenchymal cells of a patient with an iron storage disease, before and after phlebotomy. This supports the hypothesis that the process of iron unloading is an organelle specific process. Measurement of the binary morphology, represented by the area and contour ratio of the iron containing objects revealed no information about differences between the objects. This finding contradicts the visual suggestion that ferritin clusters are more irregularly shaped than the other iron objects. Also, no differences could be found in this sense between the situations before and after phlebotomy. With respect to the density appearance, objects that have an inhomogeneous iron loading averagely contain more iron. This observation does correspond well with the visual impression of the increasingly irregular appearance of more well-loaded structures.

Biopsy↗

Quantitative electron spectroscopic imaging in bio-medicine: evaluation and application.

Electron spectroscopic imaging (ESI) with the energy-filtering transmission electron microscope enables the investigation of chemical elements in ultrathin biological sections. An analysis technique has been developed to calculate elemental maps and quantitative distributions from ESI sequences. Extensive experience has been obtained with a practical implementation of this technique. A procedure for more robust element detection has been investigated and optimized. With the use of Fe-loaded Chelex beads, the measurement system has been evaluated with respect to the linearity of the element concentration scale, the reproducibility of the measurements and the visual usage of image results. In liver specimens of a patient with an iron storage disease the detectability of iron was tested and we tried to characterize iron-containing components. The concentration measurement scale is approximately linear up to a relative section thickness of approximately equal to 0.5. Monitoring of this parameter is therefore considered to be important. The reproducibility was measured in an experiment with Fe-Chelex. The iron concentration differed by 6.4% between two serial measurements. Element distributions are in many applications interpreted visually. For this purpose the frequently used net-intensity distributions are regarded as unsuitable. For the quantification and visual interpretation of concentration differences mass thickness correction has to be performed. By contrast, for the detection of elements the signal-to-noise ratio is the appropriate criterion. Application of ESI analysis demonstrated the quantitative chemical capabilities of this technique in the investigation of iron storage diseases. Based on an assumed ferritin iron loading in vivo, different iron components can be discerned in liver parenchymal cells of an iron-overloaded patient.

Ferritins↗

Estimation of fractal dimension in radiographs.

In the last decade, the fractal dimension has become a popular parameter to characterize image textures. Also in radiographs, various procedures have been used to estimate the fractal dimension. However, certain characteristics of the radiographic process, e.g., noise and blurring, interfere with the straightforward application of these estimation methods. In this study, the influence of quantum noise and image blur on several estimation methods was quantified by simulating the effect of quantum noise and the effect of modulation transfer functions, corresponding with different screen-film combinations, on computer generated fractal images. The results are extrapolated to explain the effect of film-grain noise on fractal dimension estimation. The effect of noise is that, irrespective of the noise source, the fractal dimension is overestimated, especially for lower fractal dimensions. On the other hand, blurring results in an underestimation of the dimensions. The effect of blurring is dependent on the estimation method used; the dimension estimates by the power spectrum method are lowered with a constant value, whereas the underestimation by the methods working in the spatial domain is dependent on the given dimension. The influence of the MTF and noise on fractal dimension estimation seriously limits the comparability of fractal dimensions estimated from radiographs which differ in noise content or MTF. Only when the power spectrum method is used, it is possible to correct for the influence of different MTFs of screen-film combinations. It is concluded that only when using the same object-focus distance, the same exposure conditions, the same digitizer at the same resolution, can fractal dimensions as estimated in radiographs be reliably compared.

Biophysical Phenomena↗

Energy-filtering transmission electron microscopy as a tool for structural and compositional analysis of isolated ferritin particles.

Structural and compositional analysis of isolated horse-spleen ferritin particles was performed by energy filtering transmission electron microscopy (EFTEM). Ferritin particles were collected in ultrathin (2 nm thick) chromium films and analyzed without any additional stain by electron energy-loss spectroscopy (EELS) for iron and carbon and by electron-spectroscopic imaging (ESI) for carbon. The ultrastructure of the proteinaceous shell of the ferritin particle, as obtained by the carbon net-intensity electron spectroscopical and carbon concentration-distribution images, was qualitatively compared to the structure as acquired by a negative-staining procedure. Quantitative analysis of the number of carbon atoms in the ferritin-shell proteins was carried out through an ESI-acquisition protocol and processing procedure with calibrated attenuation filters in the optical path to the TV camera. This procedure included images acquired with calibrated attenuation filters for the compensation of shading and the non-linear performance of the TV camera used in the analytical part of the procedure. A new ¿ESI-Spectra¿ program is proposed that allows element-related spectra to be generated at any place and with any frame size in a contrast-sensitive or other type of image present on the computer monitor screen.

Animals↗

Engery-filtering transmission electron microscopy of biological specimens.

By energy-filtering transmission electron microscopy (EFTEM) electrons can be separated by their energy losses. An electron-energy filter, added to the microscope column allows the measurement of the energy distribution of transmitted electrons that have lost energy (< 2,000 eV, with an energy resolution of approximately 1 eV). These filtered electrons, recorded either as a spectrum or as an image, are composed of two parts superimposed on top of each other: (a) the unspecific energy-loss population (= the continuum) and (b) the specific element-related energy-loss population (= the edges). At the edges, electron data in spectra and images are mathematically processed, to obtain the desired element-related net-intensity values or images. These data are related to the total transmitted electron intensity, from the zero- and low-loss spectral region giving the relative spectralor image intensity rations ((S)R*x, (I)R*x), which can be related to the element concentration. The acquisition of the zero-loss and low-loss data is hampered by the restricted dynamic range of the TV camera. By improvements through the introduction of calibrated attenuation filters in the optical path to the TV-camera, more reliable values for (S)R*x and (I)R*x can be acquired. By addition of Bio-standards adjacent to the tissue, a "known" and "unknown" concentration of the element present in the same ultrathin section and the "bias" in the concentration estimation, can be obtained. Some practical examples are given for the estimation of the iron cencentration in siderosomes, boron in melasosomes and calcium in calcium oxalate monohydrate crystals.

Calcium↗

Correspondence analysis for quantification in electron energy loss spectroscopy and imaging.

Electron energy loss spectroscopy (EELS) is a technique to investigate the physical properties of material. Using this technique it is possible to detect the presence of a specific element in a specimen. When used in combination with an electron microscope, energy filtered images may be obtained, which in principle may be used to quantify the local element concentration. This involves a process of background correction, conventionally performed assuming a specific parametric behavior of the spectral intensity as a function of electron energy loss. In this article a parameter-free method is described for background correction based on the formalism of correspondence analysis. Such a method may be used in parts of the spectrum where the functional dependence of the spectral intensity is unknown. Use of this method for element detection has been suggested before. This article reports simulation experiments suggesting its suitability for quantitative determination of element distributions and element concentrations.

Image Processing, Computer-Assisted↗