National Repository of Grey Literature 25 records found  1 - 10nextend  jump to record: Search took 0.03 seconds. 
Novel embedding resins for 3D-SEM II
Šlouf, Miroslav ; Pavlova, Ewa ; Strachota, Beata ; Strachota, Adam ; Nebesářová, Jana ; Týč, Jiří ; Krzyžánek, Vladislav ; Skoupý, Radim
Research report for Thermo Fisher Scientific s. r. o., based on common project awarded by TA CR (program: National center of competence, TN01000008).
Analysis of pulmonary samples infected with Aspergillus fumigatus and Pseudomonas aeruginosa by scanning electron microscopy
Juříková, Tereza ; Benada, Oldřich (advisor) ; Nebesářová, Jana (referee)
Despite the significant progress in medicine, infectious diseases are life-threatening thanks to an increasing number of multiresistant strains of microorganisms and late detection of pathological agents. An opportunistic fungus Aspergillus fumigatus cause respiratory system diseases called aspergillosis. The invasive pulmonary aspergillosis affects immunocompromised patients after inhalation of ubiquitous conidia of A. fumigatus and results in 450,000 deaths per year. The biofilm formation in the infected tissue protects A. fumigatus against antimicrobial drugs. Late therapy may not be effective. Infection of immunocompromised patients and biofilm formation is characteristic also for gram negative bacteria Pseudomonas aeruginosa, which is due to the production of many factors of virulence and multiresistance a dreaded opportunistic pathogen. Scanning electron microscopy (SEM) provides detail information about morphology of microorganisms with the resolution in range of tens of nanometers that allows to observe microorganisms in the infected tissue and its pathological changes. Mass spectrometry allows to detect infection and its course based on identification of characteristic microbial molecules. The aim of this study was to optimize sample preparation of tissues infected with A. fumigatus or P....
Electron cryo-microscopy techniques in biological research and nanotechnologies
Mistríková, Veronika ; Bednár, Jan (advisor) ; Nebesářová, Jana (referee) ; Benada, Oldřich (referee)
Preparation of biological samples for transmission electron microscopy is not a trivial task. The samples must withstand a vacuum environment present inside a microscope, and it is often necessary to use non-physiological procedures for their processing. These procedures usually involve aldehyde-based fixation, replacing water with alcohol (i.e. dehydration/substitution), and embedding into a resin, which creates support for the subsequent preparation of thin sections that can be placed into the microscope. In the last decade, the method of cryo-fixation (vitrification) using ultra-fast high-pressure freezing followed by freeze substitution and low-temperature resin embedding gained a dominant position in the cell biology research. In this way, a range of biological samples with a thicknesses up to several hundreds of micrometers was successfully vitrified to a state that was closely related to their in vivo structures. The cryo-fixation of isolated biological objects (with a limited thickness up to several micrometers) is possible in a thin layer of vitrified water by plunge freezing at ambient pressure. In combination with electron cryo-microscopy, this method has become the most effective and fundamental principle for the high-resolution studies and image analysis of fully hydrated samples...
Cathodoluminescence imaging possibilities of commercial fluorophores and cathodoluminescence probes using a functional sample of CRYTUR detector
Vancová, Marie ; Vaněček, Jiří ; Nebesářová, Jana ; Krzyžánek, Vladislav ; Skoupý, Radim
Cathodoluminescence imaging possibilities of commercial fluorophores and cathodoluminescence probes using a functional sample of CRYTUR detector
Transmission of very slow electrons as a diagnostic tool
Frank, Luděk ; Nebesářová, Jana ; Vancová, Marie ; Paták, Aleš ; Mikmeková, Eliška ; Müllerová, Ilona
The penetration of electrons through solids is retarded by sequences of their interactions with the matter in which the electron changes its direction of motion and loses its energy. Inelastic collisions, the intensity of which reaches a maximum at around 50 electronvolts (eV) and drops steeply on both sides of this fuzzy threshold, are decisive for the penetration of electrons. Transmission microscopy (TEM or STEM) observes thin samples of tens to hundreds of nanometres in thickness by passing electrons of energies of tens to hundreds of kiloelectronvolts through them. The range below 50 eV has recently been utilized in the examination of surfaces with reflected electrons, where high image resolution is achieved thanks to the retardation of electrons close to the sample surface in the ´cathode lens´ . In this lens, the role of the cathode is played by the sample itself, biased to a high negative potential. This principle can also be utilized in the transmission mode with samples of a thickness at and below 10 nm. This method has recently been implemented and verified on graphene samples prepared by various methods. The results have made it possible to diagnose the continuity and quality of the graphene flakes. Furthermore, series of experiments have been performed involving the observation of ultrathin tissue sections with electrons decelerated to about 500 eV and less, where they provide an image contrast of the cell ultrastructure much higher than that provided by traditional microscopic modes.
Analysis of pulmonary samples infected with Aspergillus fumigatus and Pseudomonas aeruginosa by scanning electron microscopy
Juříková, Tereza ; Benada, Oldřich (advisor) ; Nebesářová, Jana (referee)
Despite the significant progress in medicine, infectious diseases are life-threatening thanks to an increasing number of multiresistant strains of microorganisms and late detection of pathological agents. An opportunistic fungus Aspergillus fumigatus cause respiratory system diseases called aspergillosis. The invasive pulmonary aspergillosis affects immunocompromised patients after inhalation of ubiquitous conidia of A. fumigatus and results in 450,000 deaths per year. The biofilm formation in the infected tissue protects A. fumigatus against antimicrobial drugs. Late therapy may not be effective. Infection of immunocompromised patients and biofilm formation is characteristic also for gram negative bacteria Pseudomonas aeruginosa, which is due to the production of many factors of virulence and multiresistance a dreaded opportunistic pathogen. Scanning electron microscopy (SEM) provides detail information about morphology of microorganisms with the resolution in range of tens of nanometers that allows to observe microorganisms in the infected tissue and its pathological changes. Mass spectrometry allows to detect infection and its course based on identification of characteristic microbial molecules. The aim of this study was to optimize sample preparation of tissues infected with A. fumigatus or P....
Development of ultrastructural methods and their application in studies on the cell nucleus
Filimonenko, Anatoly ; Hozák, Pavel (advisor) ; Nebesářová, Jana (referee) ; Lanctôt, Christian (referee)
Despite the capabilities of molecular-biological methods in deciphering the interplay of different biological molecules and molecular complexes, the understanding of respective functions in living cells requires application of in situ methods. Obviously, these methods should provide maximal resolution and the best possible preservation of the biological object in a native state, as well as correct statistical evaluation of the spatial characteristics of detected molecular players. Transmission electron microscopy provides the highest possible resolution for analysis of biological samples. The simultaneous detection of biological molecules by means of indirect immunolabeling provides valuable information about their localization in cellular compartments and their possible interactions in macromolecular complexes. To analyze this, we have developed a complex stereological method for statistical evaluation of immunogold clustering and colocalization patterns of antigens on ultrathin sections, including a user-friendly interface. Functional microarchitecture of DNA replication and transcription sites has been successfully characterized using the developed stereological tools. Our data demonstrate that DNA replication is compartmentalized within cell nuclei at the level of DNA foci and support the view...
Development of surface and body musculature of the bird schistosome Trichobilharzia regenti
Bulantová, Jana ; Horák, Petr (advisor) ; Nebesářová, Jana (referee) ; Toledo, Rafael (referee)
Description of Trichobilharzia regenti as a new species of nasal bird schistosome in 1998 was only the first step in our knowledge of this extraordinary parasite. Natural definitive hosts of T. regeni are anseriform birds, but infective larvae - cercariae - are able to penetrate also into mammalian hosts including humans. There they are causative agents of hypersensitive skin immune reaction called cercarial dermatitis or swimmer's itch. Contrary to other schistosomes, miracidia of T. regenti hatch directly inside the definitive host tissue. Schistosomula migrate through the nervous system of vertebrates and, together with adult worms, they have predominantly extravascular localization in definitive hosts. Adult worms have a short lifespan and low degree of sexual dimorphism, connected with lower dependence of adult females on long-term contact with males. During the life cycle, T. regenti can be found within three different environments (freshwater, tissue of intermediate molluscan host and tissue of vertebrate host). Each of the seven developmental stages has a different role in the life cycle which corresponds with different organization of various organ systems. The introductory part of the thesis is focused entirely on ontogenetic changes of surface ultrastructure and body musculature of...

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