Možnosti využití fluorescenční mikroskopie pro stanovení toxicity nanočástic u prokaryotních mikroorganizmů
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Cílem bakalářské práce je seznámení se s problematikou stanovení toxicity nanočástic na prokaryotních mikroorganizmech. Studentka se naučí kultivaci bakterií, přípravu růstových medií a charakterizaci bakteriálních kultur. Studentka bude pracovat s nejméně třemi typy nanočástic a jejich vliv na prokaryotní buňky bude studovat pomocí nejméně dvou analytických metod, se kterými se nejprve detailně seznámí (např. stanovení ROS (Reactive Oxygen Species) v buňkách nebo určení životaschopnosti buněk (LIVE/DEAD staining). Pro srovnání výsledků použije klasickou kultivaci buňek bakterií na agarových mediích.
Abstract: The bachelor thesis discusses properties and methods for studying toxicity of different nanoparticles using prokaryotic microorganisms. Main aim of thesis was to introduce the principles and comparison of methods for determination of toxicity of nanoparticles: Determination of cell viability, Determination of Reactive Oxygen Species in cells and Cultivation of bacteria on agar media. Nanoparticles (nanodiamond, magnetite and nZVI) were characterized using the CPS disc centrifuge and Zeta Sizer (DLS and DCS methods). The toxicity was investigated on model organism of bacteria Pseudomonas putida. Selected methods gave similar results and showed that the nanoparticles are not harmful to prokaryotic P. putida. The most accurate method seems to be Determination of cell viability which describes the nanoparticles according to available literature. But each method has its advantages and disadvantages, for example Cultivation of bacteria on agar media is more time consuming. Characterization of nanoparticles provided information about the formation of large aggregates (hundreds of nm) in bacterial growth media which might influence the toxicity tests.
Abstract: The bachelor thesis discusses properties and methods for studying toxicity of different nanoparticles using prokaryotic microorganisms. Main aim of thesis was to introduce the principles and comparison of methods for determination of toxicity of nanoparticles: Determination of cell viability, Determination of Reactive Oxygen Species in cells and Cultivation of bacteria on agar media. Nanoparticles (nanodiamond, magnetite and nZVI) were characterized using the CPS disc centrifuge and Zeta Sizer (DLS and DCS methods). The toxicity was investigated on model organism of bacteria Pseudomonas putida. Selected methods gave similar results and showed that the nanoparticles are not harmful to prokaryotic P. putida. The most accurate method seems to be Determination of cell viability which describes the nanoparticles according to available literature. But each method has its advantages and disadvantages, for example Cultivation of bacteria on agar media is more time consuming. Characterization of nanoparticles provided information about the formation of large aggregates (hundreds of nm) in bacterial growth media which might influence the toxicity tests.
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Fluorescence, Reaktivní formy kyslíku, Pseudomonas putida, nanodiamant, magnetit, nZVI, nanotoxicita, Fluorescence, Reactive Oxygen Species, Pseudomonas putida, nanodiamond, magnetite, nZVI, nanotoxicity