Ekspanzijska mikroskopija u proučavanju arhitekture diobenog vretena

master's thesis

• Title (english): Expansion microscopy in exploring the architecture of the mitotic spindle
• Author: Ivana Ponjavić
• Mentor: Iva Tolić (mentor)
• Mentor: Mirjana Pavlica (mentor)
• Committee member: Mirjana Pavlica (predsjednik povjerenstva)
• Committee member: Ines Radanović (član povjerenstva)
• Committee member: Jasmina Rokov Plavec (član povjerenstva)
• Committee member: Branimir Bertoša (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Granter: University of Zagreb; Faculty of Science; (Department of Chemistry); Zagreb
• Defense date and country: 2018-10-01, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Ekspanzijska mikroskopija je nova metoda mikroskopije u kojoj se kemijskim postupcima fizički povećava fiksirani uzorak te dozvoljava da strukture bliže od granice difrakcije svjetlosti (250 nm) postanu razlučive. To je tehnika u kojoj su fluorofori na fiksiranom uzorku vezani za polimer koji ima mogućnost izotropnog ekspandiranja te kada se polimer ekspandira, omogućava super-rezolucijsku mikroskopiju sa standardnim mikroskopom. Cilj ovog rada bio je uz pomoć razvijene metode ekspanzijske mikroskopije, proučiti arhitekturu diobenog vretena, molekularne strukture odgovorne za jednaku raspodjelu kromosoma među stanicama kćerima. Vizualizirane i analizirane su RPE1 i U2OS stanice. U ovom radu razvijen je protokol za ekspanzijsku mikroskopiju, te je dobivena slika cjelovitog diobenog vretena u ljudskim stanicama uz upotrebu ekspanzijske mikroskopije. Pokazano je da snimke ekspandiranih diobenih vretena nude mnoštvo informacija o arhitekturi vretena te je u budućnosti potrebno optimizirati protokol za bilo koji protein, koji se želi vizualizirati u diobenom vretenu.

Abstract (english)

Expansion microscopy is a newly discovered microscopy technique that uses physical expansion of fixed specimens to allow features closer than the diffraction limit of light (250 nm) to become resolvable in the expanded specimen. It is a technique in which fluorophores on fixed specimens are linked to a swellable polymer that is physically expanded to enable super-resolution microscopy with ordinary microscopes. The goal of this thesis was, with the help od the developed expansion microscopy technique, to study the architecture od the mitotic spindle, a molecular machine responsible for equal distribution of chromosomes between the daughter cells. RPE1 and U2OS cells were visualized and analyzed. In this thesis the protocol for expansion micoscropy was developed and used for imaging the whole mitotic spindle in human cells. It was shown that the images of the expanded mitotic spindles offer plenty information about the architecture of the spindle and in the future it would be interesting to optimize the protocol for visualising any protein in the mitotic spindle with the expansion microscopy.

• Keywords: ekspanzijska mikroskopija; premosni i kinetohorni mikrotubuli; mitoza.
• Keywords (english): expansion microscopy; mitotic spindle; bridging fibres; mitosis
• Language: croatian
• URN:NBN: urn:nbn:hr:217:502098
• Study programme: Title: Integrated undergraduate and graduate university study of Biology and Chemistry Education; specializations in: Integrated undergraduate and graduate university study of Biology and Chemistry Education; Course: Integrated undergraduate and graduate university study of Biology and Chemistry Education; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra edukacije biologije i kemije (magistar/magistra edukacije biologije i kemije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access; Embargo expiration date: 2020-10-01
• Created on: 2018-10-29 12:04:50