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Ovrednotenje postopkov kontrastiranja bioloških vzorcev z alternativnimi kontrastnimi snovmi za presevno elektronsko mikroskopijo
ID Mahorič, Maja (Author), ID Žnidaršič, Nada (Mentor) More about this mentor... This link opens in a new window

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Abstract
Presevni elektronski mikroskop nam omogoča slikanje različnih bioloških vzorcev z visoko ločljivostjo. Ti so v splošnem sestavljeni iz elementov z nizkim atomskim številom, zato so pri slikanju slabo kontrastni. Kontrast vzorcev povečamo z uporabo kontrastnih sredstev, ki večinoma vsebujejo ione težkih kovin. V zadnjem času se na trgu pojavljajo nova kontrastna sredstva, ki poenostavijo postopek priprave vzorcev. Novejši postopki za kontrastiranje so običajno preizkušeni predvsem na različnih humanih in sesalskih vzorcih. V nalogi smo preizkusili dve novejši in redko uporabljani kontrastni sredstvi AU ZeroTM in natrijevo silicij-volframovo kislino v postopkih priprave vzorcev tkiv rib, tkiv žuželk in rastlinskih virusov za elektronsko mikroskopijo. Ovrednotili smo pridobljene mikrografije v primerjavi s klasičnimi postopki. Rezultati mikroskopiranja vzorcev jeter ribe in mišice žuželke so pokazali, da je alternativno kontrastno sredstvo AU ZeroTM v splošnem ustrezno nadomestilo klasični postopek in nam je omogočilo opazovanje značilnih struktur v vzorcu tkiva. Razlike smo opazili predvsem pri kontrastiranju heterokromatina in lipidnih kapelj v hepatocitih ribe. Rezultati kontrastiranja vzorcev v suspenziji so pokazali, da je natrijeva silicij-volframova kislina v splošnem uporabna za detekcijo in slikanje virusov mozaika tobaka paradižnika, vendar na pridobljenih slikah ultrastrukturne značilnosti virusa niso bile zelo dobro razvidne.

Language:Slovenian
Keywords:presevna elektronska mikroskopija, kontrastna sredstva, tkivne rezine, vzorci v suspenziji, priprava vzorcev za presevno elektronsko mikroskopijo
Work type:Bachelor thesis/paper
Typology:2.11 - Undergraduate Thesis
Organization:FKKT - Faculty of Chemistry and Chemical Technology
Year:2021
PID:20.500.12556/RUL-130133 This link opens in a new window
COBISS.SI-ID:82909187 This link opens in a new window
Publication date in RUL:10.09.2021
Views:2102
Downloads:126
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Secondary language

Language:English
Title:Evaluation of the procedures for biological samples contrasting with alternative contrastive agents in transmission electron microscopy
Abstract:
Transmission electron microscopy enables analyses of ultrastructural characteristics of biological samples. These samples consist mainly of elements with low atomic numbers, which results in the micrographs that lack contrast. To enhance the contrast, we use contrast agents mostly based on ions of heavy metals. The recent development of new alternative contrast agents enables an easier preparation of biological samples. New protocols are usually tested on different human and mammal samples. In these study, we used two new and rarely used contrast agents AU ZeroTM and sodium silicotungstate for preparation of fish tissue, insect tissue and plant viruses for transmission electron microscopy. We compared and evaluated the acquired micrographs with respect to conventional protocols. The results of imaging the ultrathin sections of fish liver and of insect muscle showed that agent AU ZeroTM generally adequately contrasted these samples and it enabled us to resolve the ultrastructure of our samples. Differences were observed in the case of heterochromatin and lipid droplets imaging in fish hepatocytes. The micrographs of samples in suspension showed that sodium silicotungstate is generally appropriate for detection and observation of tomato mosaic virus but did not enable a more precise observation of the virus’s ultrastructural characteristics.

Keywords:transmission electron microscopy, contrast agents, tissue samples, samples in suspension, sample preparation for transmission electron microscopy

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