low-energy electron microscopy
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... The basic principles of the electron microscope 1 -- 1.1 The use of electrons for microscopy 2 -- 1.2 ... ... Electron lenses 4 -- 1.3 Defects of electron lenses 9 -- 1.3.1 Spherical aberration 9 -- 1.3.2 Chromatic ... ... The design of the electron microscope 38 -- 2.1 General construction of the column 38 -- 2.2 The electron ... ... 313 -- 9.3.3 Energy analysis 316 -- 9.3.4 Scanning electron diffraction 319 -- 9.4 Scanning transmission ... ... electron microscopy 320 -- 9.4.1 The scanning transmission electron microscope (STEM) 322 -- 9.4.2 The ...
Practical methods in electron microscopy ; v. 2
Laboratory edition xii, 345 stran : ilustrace ; 23 cm
- MeSH
- elektronová mikroskopie MeSH
- fyzika MeSH
- mikroskopie MeSH
- Publikační typ
- monografie MeSH
- Konspekt
- Fyzika
- NLK Obory
- fyzika, biofyzika
Leachates collected at two (active and closed) municipal solid waste (MSW) landfills were examined for colloids and particles by transmission electron microscopy, energy dispersive spectrometry, selected area electron diffraction and for the chemical compositions of the filtrates after the filtration to 0.1 µm and ultrafiltration to 1 kDa (~ 1 nm). Six groups of colloids/particles in the range 5 nm to 5 µm were determined (in decreasing order of abundance): carbonates, phyllosilicates (clay minerals and micas), quartz, Fe-oxides, organics and others (salts, phosphates). Inorganic colloids/particles in leachates from the active landfill predominantly consist of calcite (CaCO(3)) and minor clay minerals and quartz (SiO(2)). The colloids/particles in the leachates from the closed landfill consist of all the observed groups with dominant phyllosilicates. Whereas calcite, Fe-oxides and phosphates can precipitate directly from the leachates, phyllosilicates and quartz are more probably either derived from the waste or formed by erosion of the geological environment of the landfill. Low amounts of organic colloids/particles were observed, indicating the predominance of organic molecules in the 'truly dissolved' fraction (fulvic compounds). Especially newly formed calcite colloids forming particles of 500 nm and stacking in larger aggregates can bind trace inorganic contaminants (metals/metalloids) and immobilize them in landfill environments.
The limited availability of biological samples hinders phylogenetic efforts to define structural differences among various biological groups. A novel workflow enabling the analysis of protists in low cell numbers by electron microscopy (EM) is described with cysts of Giardia intestinalis, a single-celled eukaryotic parasite. Correlative light and electron microscopy (CLEM) allows for the selection of individual cells and is economical in terms of time and cost. We describe a cyst purification protocol in combination with an adhesive coating for fixation and ultrathin embedding that results in excellent preservation of cell morphology. The application of advanced structural and analytical EM methods, such as high-resolution field emission scanning electron microscopy (FESEM), focused ion beam tomography (FIB/SEM), and energy-dispersive X-ray spectroscopy (EDX) analysis, is demonstrated. The workflow represents a new approach for studying the cellular and organelle architecture of rare and "difficult to culture" microorganisms.
The Pithoviridae giant virus family exhibits the largest viral particle known so far, a prolate spheroid up to 2.5 μm in length and 0.9 μm in diameter. These particles show significant variations in size. Little is known about the structure of the intact virion due to technical limitations with conventional electron cryo-microscopy (cryo-EM) when imaging thick specimens. Here we present the intact structure of the giant Pithovirus sibericum particle at near native conditions using high-voltage electron cryo-tomography (cryo-ET) and energy-filtered cryo-EM. We detected a previously undescribed low-density outer layer covering the tegument and a periodical structuring of the fibres in the striated apical cork. Energy-filtered Zernike phase-contrast cryo-EM images show distinct substructures inside the particles, implicating an internal compartmentalisation. The density of the interior volume of Pithovirus particles is three quarters lower than that of the Mimivirus. However, it is remarkably high given that the 600 kbp Pithovirus genome is only half the size of the Mimivirus genome and is packaged in a volume up to 100 times larger. These observations suggest that the interior is densely packed with macromolecules in addition to the genomic nucleic acid.
Although myriads of experimental approaches have been published in the field of fungal infection diagnostics, interestingly, in 21st century there is no satisfactory early noninvasive tool for Aspergillus diagnostics with good sensitivity and specificity. In this work, we for the first time described the fungal burden in rat lungs by multimodal imaging approach. The Aspergillus infection was monitored by positron emission tomography and light microscopy employing modified Grocott's methenamine silver staining and eosin counterstaining. Laser ablation inductively coupled plasma mass spectrometry imaging has revealed a dramatic iron increase in fungi-affected areas, which can be presumably attributed to microbial siderophores. Quantitative elemental data were inferred from matrix-matched standards prepared from rat lungs. The iron, silver, and gold MS images collected with variable laser foci revealed that particularly silver or gold can be used as excellent elements useful for sensitively tracking the Aspergillus infection. The limit of detection was determined for both (107) Ag and (197) Au as 0.03 μg/g (5 μm laser focus). The selective incorporation of (107) Ag and (197) Au into fungal cell bodies and low background noise from both elements were confirmed by energy dispersive X-ray scattering utilizing the submicron lateral resolving power of scanning electron microscopy. The low limits of detection and quantitation of both gold and silver make ICP-MS imaging monitoring a viable alternative to standard optical evaluation used in current clinical settings.
- MeSH
- Aspergillus izolace a purifikace patogenita MeSH
- aspergilóza diagnóza diagnostické zobrazování mikrobiologie MeSH
- krysa rodu rattus MeSH
- lidé MeSH
- mikroskopie elektronová rastrovací metody MeSH
- multimodální zobrazování metody MeSH
- plíce diagnostické zobrazování mikrobiologie patologie MeSH
- pozitronová emisní tomografie metody MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Quantum dots (QDs) absorb ultraviolet and long-wavelength visible light energy much more efficiently than natural bacterial light-harvesting proteins and can transfer the excitation energy to photosynthetic reaction centers (RCs). Inclusion of RCs combined with QDs as antennae into liposomes opens new opportunities for using such hybrid systems as a basis for artificial energy-transforming devices that potentially can operate with greater efficiency and stability than devices based only on biological components or inorganic components alone. RCs from Rhodobacter sphaeroides and QDs (CdSe/ZnS with hydrophilic covering) were embedded in lecithin liposomes by extrusion of a solution of multilayer lipid vesicles through a polycarbonate membrane or by dialysis of lipids and proteins dispersed with excess detergent. The efficiency of RC and QD interaction within the liposomes was estimated using fluorescence excitation spectra of the photoactive bacteriochlorophyll of the RCs and by measuring the fluorescence decay kinetics of the QDs. The functional activity of the RCs in hybrid complexes was fully maintained, and their stability was even increased. The efficiency of energy transfer between QDs and RCs and conditions of long-term stability of function of such hybrid complexes in film preparations were investigated as well. It was found that dry films containing RCs and QDs, maintained at atmospheric humidity, are capable of maintaining their functional activity for at least some months as judged by measurements of their spectral characteristics, efficiency of energy transfer from QDs to RCs and RC electron transport activity. Addition of trehalose to the films increases the stability further, especially for films maintained at low humidity. These stable hybrid film structures are promising for further studies towards developing new phototransformation devices for biotechnological applications.
BACKGROUND: Because dacryoliths occur at low frequency, few studies have focused on their composition. We aimed to present findings from morphological, chemical, and mineralogic analysis of 86 dacryoliths. METHODS: We studied 86 dacryoliths obtained during 832 dacryocystorhinostomies (DCR) performed for postsaccal obstruction. We examined the samples with atomic infrared spectrometry (80 samples), amino acid analysis (17 samples), scanning electron microscopy, and an electron microprobe with an energy dispersive detector (seven samples). RESULTS: Dacryoliths were found in 86/832 DCRs (10.3 %), mostly in patients with primary acquired nasolacrimal duct obstruction. All the dacryoliths were soft, composed of organic material, including proteins and mucoproteins, with approximately 20 % amino acid content. There were no "hard" dacryoliths composed of calcium phosphate. The stones were composed of lobes and lobules built on an amorphous core material with small cavities, probably as a result of various chemical processes that produced a gaseous product. The most frequent elements found in inorganic inclusions were silicon, magnesium, sulfur, potassium, calcium, sodium, and chlorine. Also, some particles had high contents of bismuth, titanium, iron, and organic fibers. The fibers found in the core of dacryoliths suggested a potential origin from cotton swabs used in cosmetics. CONCLUSION: Dacryoliths are composed almost exclusively of organic material, including proteins and mucoproteins, with approximately 20 % amino acid content.
- MeSH
- aminokyseliny analýza MeSH
- dakryocystorinostomie MeSH
- dítě MeSH
- dospělí MeSH
- ductus nasolacrimalis patologie MeSH
- kaménky chemie ultrastruktura MeSH
- kojenec MeSH
- lidé středního věku MeSH
- lidé MeSH
- litiáza chemie diagnóza chirurgie MeSH
- mikroanalýza elektronovou sondou MeSH
- mikroskopie elektronová rastrovací MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mukoproteiny analýza MeSH
- nemoci slzného ústrojí diagnóza chirurgie MeSH
- předškolní dítě MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- spektroskopie infračervená s Fourierovou transformací MeSH
- vápník analýza MeSH
- Check Tag
- dítě MeSH
- dospělí MeSH
- kojenec MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- předškolní dítě MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Low voltage electron microscopes working in transmission mode, like LVEM5 (Delong Instruments, Czech Republic) working at accelerating voltage 5 kV or scanning electron microscope working in transmission mode with accelerating voltage below 1 kV, require ultrathin sections with the thickness below 20 nm. Decreasing of the primary electron energy leads to enhancement of image contrast, which is especially useful in the case of biological samples composed of elements with low atomic numbers. As a result treatments with heavy metals, like post-fixation with osmium tetroxide or ultrathin section staining, can by omitted. The disadvantage is reduced penetration ability of incident electrons influencing the usable thickness of the specimen resulting in the need of ultrathin sections of under 20 nm thickness. In this study we want to answer basic questions concerning the cutting of extremely ultrathin sections: Is it possible routinely and reproducibly to cut extremely thin sections of biological specimens embedded in commonly used resins with contemporary ultramicrotome techniques and under what conditions? Microsc. Res. Tech. 79:512-517, 2016. © 2016 Wiley Periodicals, Inc.
- MeSH
- design vybavení MeSH
- elektronová mikroskopie přístrojové vybavení metody MeSH
- epoxidové pryskyřice chemie MeSH
- mikrotomie metody MeSH
- myokard ultrastruktura MeSH
- myši MeSH
- polymery chemie MeSH
- srdce diagnostické zobrazování MeSH
- zalévání tkání plastickou hmotou metody MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
