Weighted simultaneous iterative reconstruction technique for single-axis tomography.

Tomographic techniques play a crucial role in imaging methods such as transmission electron microscopy (TEM) due to their unique capabilities to reconstruct three-dimensional object information. However, the accuracy of the two standard tomographic reconstruction techniques, the weighted back-projection (W-BP) and the simultaneous iterative reconstruction technique (SIRT) is reduced under common experimental restrictions, such as limited tilt range or noise. We demonstrate that the combination of W-BP and SIRT leads to an improved tomographic reconstruction technique: the weighted SIRT. Convergence, resolution and reconstruction error of the W-SIRT are analyzed by a detailed analytical, numerical, and experimental comparison with established methods. Our reconstruction technique is not restricted to TEM tomography but can be applied to all problems sharing single axis imaging geometry.

Electron holography of biological samples.

In this paper, we summarise the development of off-axis electron holography on biological samples starting in 1986 with the first results on ferritin from the group of Tonomura. In the middle of the 1990s strong interest was evoked, but then stagnation took place because the results obtained at that stage did not reach the contrast and the resolution achieved by conventional electron microscopy. To date, there exist only a few ( approximately 12) publications on electron holography of biological objects, thus this topic is quite small and concise. The reason for this could be that holography is mostly established in materials science by physicists. Therefore, applications for off-axis holography were powerfully pushed forward in the area of imaging, e.g. electric or magnetic micro- and nanofields. Unstained biological systems investigated by means of off-axis electron holography up to now are ferritin, tobacco mosaic virus, a bacterial flagellum, T5 bacteriophage virus, hexagonal packed intermediate layer of bacteria and the Semliki Forest virus. New results of the authors on collagen fibres and surface layer of bacteria, the so-called S-layer 2D crystal lattice are presented in this review. For the sake of completeness, we will shortly discuss in-line holography of biological samples and off-axis holography of materials related to biological systems, such as biomaterial composites or magnetotactic bacteria.

Inelastic electron holography as a variant of the Feynman thought experiment.

Using a combination of electron holography and energy filtering, interference fringes produced after inelastic interaction of electrons with hydrogen molecules are examined. Surprisingly, the coherence of inelastic scattering increases when moving from the surface of a hydrogen-containing bubble to the vacuum. This phenomenon can be understood in terms of the Feynman two-slit thought experiment with a variable ambiguity of the which-way registration.

Experiments on inelastic electron holography.

Using the combination of an electron biprism and an energy filter, the coherence distribution in an inelastically scattered wave-field is measured. It is found that the degree of coherence decreases rapidly with increasing distance between two superimposed points in the object, and with increasing energy-loss. In a Si sample, coherence of plasmon scattering increases in vacuum with the distance from the edge of the sample.

Measurement of crystal thickness and orientation from selected-area Fourier transformation of a high-resolution electron hologram.

Precise knowledge of crystal thickness and orientation is critical for reliable interpretation of high-resolution transmission electron micrographs. In this paper, we propose a criterion of S(2)(T, u, v), which measures the crystal thickness by intensity matching of the selected-area Fourier transform of experimental holograms with the calculated electron diffraction pattern at a series of trial thicknesses (T) and crystal tilts (u, v). This criterion has been demonstrated successfully for local thickness determination from a simulated high-resolution image of a wedge-shaped YBa(2)Cu(3)O(7-delta) and from an experimental hologram of a Si crystal.

Plasmon holographic experiments: theoretical framework.

A theoretical framework is described to understand the results of plasmon holography experiments leading to insight in the meaning of the experimental results and pointing out directions for future experiments. The framework is based on the formalism of mutual intensity to describe how coherence is transferred through an optical system. For the inelastic interaction with the object, an expression for the volume plasmon excitations in a free electron gas is used as a model for the behaviour of aluminium. The formalism leads to a clear graphical intuitive tool for understanding the experiments. It becomes evident that the measured coherence is solely related to the angular distribution of the plasmon scattering in the case of bulk plasmons. After describing the framework, the special case of coherence outside a spherical particle is treated and the seemingly controversial idea of a plasmon with a limited coherence length obtained from experiments is clarified.

Electron holography of non-stained bacterial surface layer proteins.

We report transmission electron microscopy (TEM) investigations on bacterial surface layers (S-layers) which belong to the simplest biomembranes existing in nature. S-layers are regular 2D protein crystals composed of single protein or glycoprotein species. In their native form, S-layers are weak phase objects giving only poor contrast in conventional TEM. Therefore, they are usually examined negatively stained. However, staining with heavy metal compounds may cause the formation of structural artefacts. In this work, electron microscopy studies of non-stained S-layers of Bacillus sphaericus NCTC 9602 were performed. Compared to other proteins, these S-layers are found relatively stable against radiation damage. Electron holography was applied where information about phase and amplitude of the diffracted electron wave is simultaneously obtained. In spite of small phase shifts observed, the phase image reconstructed from the hologram of the non-stained S-layer is found to be sensitive to rather slight structure and thickness variations. The lateral resolution, obtained so far, is less than that of conventional electron microscopy of negatively stained S-layers. It corresponds to the main lattice planes of 12.4 nm observed in the reconstructed electron phase image. In addition, as a unique feature of electron holography the phase image provides thickness information. Thus, the existence of double layers of the protein crystals could be easily visualized by the height profile of the specimen.

Correction of aberration for a high-resolution electron hologram by means of the amplitude contrast criterion of image wave.

In order to further improve the resolution for a high-resolution electron hologram, the aberration working on the hologram must be corrected. Since it is rather difficult to precisely control aberration coefficients in the experimental stage, we proposed an amplitude contrast D criterion of imaging wave to determine the working aberration from the hologram itself. In the determination or correction of the aberration, we assume a symmetrical aberration function is parameterized only by a spherical aberration coefficient and a defocus value. First, D is calculated from a holographically reconstructed imaging wave of YBa(2)Cu(3)O(7-x) for each combination of these parameters. The working aberration on the imaging wave is determined from the combination of the parameters by noting the maximum or minimum D of the imaging wave at some specifically chosen thickness regions. The theoretical validity for the D criterion is then proved with three-beam dynamical diffraction formula. Finally, the 'experimental' examination for the D criterion is successfully performed on the reconstructed image wave for the Sigma=9 interface structure of a wedge-shaped silicon sample.

Inelastic electron holography

Coherence of inelastically scattered electrons was investigated by means of biprism interference experiments performed in a transmission electron microscope equipped with a highly coherent field emission gun and an imaging filter. The experimental results show that within the wave inelastically scattered at aluminium plasmons there is in fact an area of about 10 nm diameter with coherence sufficient to take electron holograms.

Inelastic scattering and holography

The controversy about whether or not an inelastically scattered electron wave can still interfere with a reference wave is solved by treating the whole problem rigorously and describing electron, source and object in one Hamiltonian. It turns out that, in principle, interference can occur between an inelastically scattered wave and a reference wave from the incident beam spectrum provided the energy difference is smaller than about 10(-15) eV. However, it is argued that the density of states in source object and electron wave is much too small to make this effect observable.

[Diagnostic pneumology using model aerosols]. / Pneumologische Diagnostik mit Modellaerosolen.

Clearance of Aerosol Particles from the Airways: Measurement of the efficiency and kinetics of particle clearance from the airways is dependent on the site of particle deposition within the lungs. Insoluble particles deposited in the alveolar region are mainly cleared by macrophages over a period of hundreds of days, whereas particles from the tracheobronchial airways are mainly cleared within hours or days by mucociliary clearance. We introduce a method for mucociliary clearance measurements called 'Radio-Bolus-Scintigraphy'. A small volume of radiolabelled aerosol particles sandwiched in clean air (aerosol bolus) is inhaled near the end of a tidal breath which leads to a preferential deposition of the particles in the airways. Particles were Fe3O4 labelled with 99mTc with an aerodynamic particle diameter of 3.5 microns (monodisperse). The retention of the particles within the lungs was detected by Gamma Camera and Human Scintillation Counter. 26 healthy subjects volunteered in this study (14 nonsmokers and 12 smokers). The half width of the mucociliary clearance was found to be 2.4 h in nonsmokers and 3.3 h in smokers (p < 0.05). Intersubject variability was small, 24 hours after inhalation 55% (+/- 6%) of the inhaled particles were still found in the airways of the subjects. No difference was found between nonsmokers and smokers. Good intersubject reproducibility makes this method useful in therapy control as well as in early diagnosis of changes in mucociliary clearance kinetics and efficiency.

Imaging of lung cancer with fluorine-18 fluorodeoxyglucose: comparison of a dual-head gamma camera in coincidence mode with a full-ring positron emission tomography system.

Dual-head gamma cameras operated in coincidence mode are a new approach for tumour imaging using fluorine-18 fluorodeoxyglucose (FDG). The aim of this study was to assess the diagnostic accuracy of such a camera system in comparison with a full-ring positron emission tomography (PET) system in patients with lung cancer. Twenty-seven patients (1 female, 26 males, age 62+/-9 years) with lung cancer or indeterminate pulmonary nodules were studied on the same day with a full-ring PET scanner (Siemens ECAT EXACT) and a coincidence gamma camera system (ADAC Vertex MCD). Sixty minutes after injection of 185-370 MBq FDG, a scan of the chest was performed with the full-ring system. Approximately 2 h p.i., the coincidence camera study was performed. Coincidence gamma camera (CGC) and PET images with (PETac) and without attenuation correction (PETnac) were analysed independently by two blinded observers. In addition, FDG uptake in primary tumours and involved lymph nodes was quantified relative to normal contralateral lung (T/L ratios). All primary tumours were histologically proven. The lymph node status was histologically determined in 23 patients. In four patients, no lymph node sampling was performed because of extensive disease or concurrent illnesses. In the 27 patients, 25 primary lung cancers and two metastatic lesions were histologically diagnosed. The number of coincidences per centimetre axial field of view was 3.33+/-0. 93x10(5) for the CGC and 1.09+/-0.36x10(6) for the dedicated PET system. All primary tumours (size: 4.6+/-2.6 cm) were correctly identified in the CGC and dedicated PET studies. T/L ratios were 4. 7+/-2.5 for CGC and 6.9+/-2.8 for PETnac (P <0.001). Histopathological evaluation revealed lymph node metastases in 11 of 88 sampled lymph node stations (size: 2.3+/-1.0 cm). All lymph node metastases were identified in the PETac studies, while PETnac detected 10/11 and CGC 8/11. For positive lymph nodes that were visible in CGC and PETnac studies, T/L ratios were 3.7+/-2.3 for CGC and 6.6+/-3.1 for PETnac (P=0.02). The diameters of false-negative lymph nodes in the CGC studies were 0.75, 1.5 and 2 cm. False-positive FDG uptake in lymph nodes was found in two patients with all three imaging methods. For all lesions combined, T/L ratios in CGC relative to PETnac studies decreased significantly with decreasing lesion size (r=0.62; P<0.001). In conclusion, compared with a full-ring PET system the sensitivity of CGC imaging for detection of lung cancer is limited by a lower image contrast which deteriorates with decreasing lesion size. Nevertheless, the ability of CGC imaging to detect pulmonary lesions with a diameter of at least 2 cm appears to be similar to that of a full-ring system. Both systems provide a similar specificity for the evaluation of lymph node involvement.

Reflection electron microscopy and interferometry of atomic steps on gold and platinum single crystal surfaces.

Two beam interferences produced using an electrostatic biprism, which is inserted in the position of the selected area diaphragm of a commercial electron microscope, may be used in reflection electron microscopy to determine the phase shifts induced by structures on single crystal surfaces. A description of our interferometrical and holographical experiments on the phase shift at steps on (111)Au and (111)Pt single crystal surfaces is given and a straight forward interpretation of the results in terms of refraction will be discussed. As a particular result phase shifts of pi and 0.9 pi were measured for monatomic steps on (111)gold and (111)platinum surfaces, respectively.

[Scintigraphic studies of pulmonary ventilation]. / Szintigraphische Untersuchungen zur Lungenbelüftung.

Nuclear medical imaging of the lung after inhalation of Tc-labelled aerosols can be used instead of ventilation scintigraphy with Xe-133 in those cases, where the single-breath-period is essential for diagnosis. Especially for pulmonary embolism aerosols have some advantages compared with Xe-133: Scintigraphic images can be done in several views, aerosols are always available, there is no problem with room-contamination. In patients with obstructive lung disease or emphysema, where the wash-out-period is more essential for diagnosis than the single-breath-period, there is an advantage of xenon to aerosols. Additionally aerosols can be used to study mucociliary function.

[Determination of left ventricular function using nuclear medicine methods]. / Beurteilung der Funktion des linken Ventrikels mit nuklearmedizinischen Methoden.

ECG-gated blood-pool scintigraphy with multiple gating acquisition is a good tool for evaluation of global and regional ventricular function. During exercise there is an average increase of ejection fraction of about 13.4% compared with rest ejection fraction in patients with normal ventricular function. A decrease or insufficient increase of ejection fraction by exercise is due to myocardial ischemia. After administration of isosorbiddinitrat in retard preparation there is an increase of ejection fraction of about 7%, lasting at least for 12 hours.

[Myocardial scintigraphy using 201 thallium]. / Myokardszintigraphie mit 201Thallium

Myocardial scintigraphy with 201Thallium is a non-invasive method for detection of myocardial infarction and coronary heart disease. Redistribution-analysis as a sequential-scintigraphy of an exercise-scan permits to distinguish between myocardial scars and coronary vessel disease.