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X-ray computed tomography of cartilage tissue of embryos
Tesařová, Markéta ; Křivánek,, Jan (referee) ; Zikmund, Tomáš (advisor)
Research of the cartilage is standardly realized by optical microscopy on histological slices. However, for better understanding of shape-organizing processes, imaging in 3D is required. This thesis deals with X-ray computed tomography of mouse embryos. Specifically, it focuses on visualization of craniofacial cartilage. The thesis summarizes the basic principles of X-ray computed tomography. Short research study dealing with staining of soft tissues for X-ray is also provided. The procedure of implementation of the tomographic measurement of mouse embryos stained by PTA is described. This procedure includes the staining protocol, fixing the sample and setting of the CT machine. Subsequent segmentation and creating of a 3D model of cartilage is also desribed. Results for every step of this procedure are demonstrated at the mouse embryos of developmental stage from 12.5 to 17.5 days. In order to improve data quality and standardization of the whole process, parameters of X-ray tube and temperature conditions during the tomographic measurement are explored. Finally, resulting models of craniofacial cartilage are shown with the possibility of subsequent analysis.
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Porovnání žlutých tělísek v pozdní embryonální fázi u primiparních a multiparních dojnic
Matoušková, Jana
Thesis has been focused on the development of corpus luteum and embryos in Holstein cows. Corpus luteum and embryos was detected and measured by ultrasonography. The aim of thesis was determine the effect of corpus luteum and other factors on the size of embryos. Results showed that the size of corpus luteum has no influence on the size of embryos. This research area is still scarce and further experiments will be needed to deepen understanding of the factors affecting embryo development.
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Abiotické a biotické faktory ovlivňující klíčení rostlin
Berka, Miroslav
Seed germination and early plant development is a crucial phase of plants' life. Multiple internal and external stimuli influence germination progress and have a serious impact on a plant's survival and vitality. Biotic and abiotic stimuli trigger a whole range of changes, both on molecular and developmental levels, but the complex molecular mechanisms regulating these responses are far from being resolved. This thesis reviews the seed germination process and outlines the role of external stimuli in its progress. The experimental part describes the development of a method for seed germination monitoring, provides new insight into the role of hydrogen peroxide in germination, and analyzes effects of cadmium ions, temperature, salt and drought on proteome and metabolome of germinating seeds of Hordeum vulgare. In total, 2000 proteins and 800 metabolites were identified. The analyses revealed over 95 putative abiotic stress markers, including 63 and 36 proteins and metabolites, respectively
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X-ray computed tomography of cartilage tissue of embryos
Tesařová, Markéta ; Křivánek,, Jan (referee) ; Zikmund, Tomáš (advisor)
Research of the cartilage is standardly realized by optical microscopy on histological slices. However, for better understanding of shape-organizing processes, imaging in 3D is required. This thesis deals with X-ray computed tomography of mouse embryos. Specifically, it focuses on visualization of craniofacial cartilage. The thesis summarizes the basic principles of X-ray computed tomography. Short research study dealing with staining of soft tissues for X-ray is also provided. The procedure of implementation of the tomographic measurement of mouse embryos stained by PTA is described. This procedure includes the staining protocol, fixing the sample and setting of the CT machine. Subsequent segmentation and creating of a 3D model of cartilage is also desribed. Results for every step of this procedure are demonstrated at the mouse embryos of developmental stage from 12.5 to 17.5 days. In order to improve data quality and standardization of the whole process, parameters of X-ray tube and temperature conditions during the tomographic measurement are explored. Finally, resulting models of craniofacial cartilage are shown with the possibility of subsequent analysis.
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Resistance of some species of fish embryos to cryoprotectants at low temperatures
ALDORF, Milan
Different cryoprotectant media for cryopreservation of embryos has been tested on model species, i.e. common carp (Cyprinus carpio) and common tench (Tinca tinca). The aim of the study was to obtain such cryoprotectants, which will be acceptable for freezing embryos up to the temperature {--}196 oC. Cryoprotectants of 10 % and 20 % methanol or 10 % and 20 % glycerin have been tested on the tench for 21 minutes of incubation on embryos of four stages, meaning at 11, 17, 23 and 29-hrs after activation of gametes. The results showed that the tench embryos were most resistant either to low temperature and or to the application of cryoprotectants in the stage of 29-hrs post gametes activation. On the other hand lower resistances were obtained in the stage of 11-hrs post gamete activation. Embryos of carp 2, 6, 22, 24 and 42-hrs after gametes activation at temperature 18 and 22 oC have been used for testing of concentration series of cryoprotectant methanol and two solutions marked VS1 and VS2 after previous disruption of egg envelope in enzyme alcalaze solution. Results showed linear decreasing resistance of embryos depending on increasing concentration of cryoprotectant methanol. Hatching success even at highest concentration of solution VS1 and VS2 has not declined below 70 %. Achieved results with solution VS2 have been subsequently used for freezing of carp embryos by special methods in cryobiology {--} vitrification. First results showed up to 4 % success of survival after freezing of embryos at {$-$}196 oC.
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