| Summary: | Submitted by Jos? Henrique Henrique (jose.neves@ufvjm.edu.br) on 2018-02-07T18:45:29Z
No. of bitstreams: 2
license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)
camila_cristina_rodrigues.pdf: 1879433 bytes, checksum: 62f0da52d006b25928023dcba20ee6c0 (MD5) === Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2018-03-09T19:14:07Z (GMT) No. of bitstreams: 2
license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)
camila_cristina_rodrigues.pdf: 1879433 bytes, checksum: 62f0da52d006b25928023dcba20ee6c0 (MD5) === Made available in DSpace on 2018-03-09T19:14:07Z (GMT). No. of bitstreams: 2
license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)
camila_cristina_rodrigues.pdf: 1879433 bytes, checksum: 62f0da52d006b25928023dcba20ee6c0 (MD5)
Previous issue date: 2017 === As leveduras do g?nero Saccharomyces, em especial as da esp?cie Saccharomyces cerevisiae, s?o microrganismos eucari?ticos unicelulares do Reino Fungi. Possuem um papel importante em processos de fermenta??o, como na utiliza??o para a produ??o de bebidas alco?licas e p?es, sendo considerados micro-organismos seguros para alimentos. O monitoramento das mat?rias-primas e da prolifera??o de leveduras ? uma necessidade para o sucesso do processo fermentativo. Nesse sentido, a citometria de fluxo, que ? um m?todo r?pido e preciso, pode se apresentar como ferramenta promissora no campo da fermenta??o e no controle de bioprocessos. Assim, o objetivo desse trabalho foi realizar a an?lise do crescimento e heterogeneidade populacional de uma linhagem comercial de Saccharomyces cerevisiae pela t?cnica de citometria de fluxo. Para isso foi utilizada a marca??o intracelular com o fluorocromo ?ster succinimid?lico de diacetato de carboxifluoresce?na. A suspens?o de levedura incorporada com o fluorocromo foi transferida para tubos de ensaio contendo 10 mL de YEPD e incubada a 25? C e foram analisadas, ap?s diferentes tempos, por citometria de fluxo, microscopia confocal e espectrofotometria. Os resultados demonstraram que foi poss?vel observar, por citometria de fluxo, altera??es morfol?gicas da linhagem Saccharomyces cerevisiae ao longo do tempo, bem como quantificar o decaimento da fluoresc?ncia do corante CFSE ap?s sucessivos processos mit?ticos. Por essa t?cnica tamb?m foi poss?vel mensurar o n?mero de c?lulas da linhagem estudada ao longo do processo de multiplica??o comparando-se ?s outras metodologias comumente utilizadas para este fim. Conclui-se com esse estudo que a citometria de fluxo mostrou-se como uma ferramenta confi?vel e sens?vel para a an?lise do crescimento e heterogeneidade populacional de Saccharomyces cerevisiae. === Disserta??o (Mestrado) ? Programa de P?s-gradua??o em Ci?ncias Farmac?uticas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2017. === Yeasts of the genus Saccharomyces, especially those of the species Saccharomyces cerevisiae, are unicellular eukaryotic microorganisms of the Fungi Kingdom. They have an important role in fermentation processes, such as in the use for the production of alcoholic beverages and breads, being considered microorganisms safe for food. The monitoring of raw materials and the proliferation of yeasts is a necessity for the success of the fermentation process. In this sense, flow cytometry, which is a fast and precise method, can be presented as a promising tool in the field of fermentation and in the control of bioprocesses. Thus, the objective of this work was to analyze the growth and population heterogeneity of a commercial strain of Saccharomyces cerevisiae by the flow cytometry technique. For this, intracellular labeling was used with the fluorochrome succinimidyl ester of carboxyfluorescein diacetate. The yeast suspension incorporated with the fluorochrome was transferred into test tubes containing 10 ml of YEPD and incubated at 25 ? C and analyzed after different times by flow cytometry, confocal microscopy and spectrophotometry. The results demonstrated that it was possible to observe, by flow cytometry, morphological alterations of the Saccharomyces cerevisiae lineage over time, as well as to quantify the decay of CFSE dye fluorescence after successive mitotic processes. By this technique it was also possible to measure the number of cells of the lineage studied throughout the multiplication process, comparing to the other methodologies commonly used for this purpose. It is concluded with this study that flow cytometry proved to be a reliable and sensitive tool for the analysis of the growth and population heterogeneity of Saccharomyces cerevisiae.
|
|---|
