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[pt] As nanopartículas magnéticas (NPMs) apresentam grande potencial em diversas aplicações tecnológicas e vem ganhando destaque na área da biomedicina devido suas propriedades superparamagnéticas. Para este trabalho foram sintetizadas nanopartículas de óxido de ferro (Fe3O4) pelo método de coprecipitação e recobertas com Pluronic F-127 (PL F-127), o qual se demonstrou, a partir de estudos anteriores, um surfactante com ótima estabilidade coloidal e alto grau de biocompatibilidade, sendo tais características relevantes para as aplicações na área da biomedicina. O método de produção se mostrou eficiente para produção de uma grande quantidade de amostra e baixo grau de oxidação, o que mantém a integridade dos resultados e do material para diversas análises no decorrer de longos períodos. A espectroscopia Raman e a difração de elétrons apontam para a composição majoritária de magnetita cristalina das amostras. As imagens obtidas através de microscopia de transmissão (MET) mostraram que o diâmetro médio das NPMs não é afetado pela concentração PL F-127 e está de acordo com os tamanhos obtidos pelas técnicas magnéticas. O MET também mostrou partículas monodispersas com formato esféricas. As técnicas de microscopia magnética de varredura (MMV), magnetômetro de amostra vibrante e de efeito Hall revelaram que o comportamento das NPMs é superparamagnético em temperatura ambiente e que a funcionalização não interferiu significativamente na magnetização de saturação. / [en] Magnetic nanoparticles (MNPs) have great potential in several technological applications and are gaining prominence in the biomedical area due to their superparamagnetic properties. For this work, iron oxide (Fe3O4) nanoparticles were synthesized by the coprecipitation method and coated with Pluronic F-127 (PL F127), which was demonstrated, from previus studies, a surfactante with excellent coloidal stability and high degree of biocompatibility, such characteristics being relevant for applications in the área of biomedicine. The production method proved to be efficient for producing a large amount of sample and a low degree of oxidation, which maintains the integrity of the results and material for several analyzes over long periods. Raman spectroscopy and eléctron diffraction indicate the samples are pure and crystalline magnetite. The images obtained through transmission eléctron microscopy (TEM) showed that the mean diameter of MNPs is not affected by the PL F-127 concentration and is in agreement with the sizes obtained by magnetic techniques. TEM also showed monodisperse particles with a spherical shape. Scanning magnetic microscopy (MMV), vibrating sample magnetometer and Hall effect techniques revealed that the behavior of NPMs is superparamagnetic at ambient temperature and that the functionalization did not significantly interfere in the saturation magnetization.

A key component of biological research is cell culture technology, which allows researchersto examine the behavior and functionality of cells in controlled environments. Conventionalcell culture monitoring frequently necessitates taking the cultures out of their incubators tomake observations under a microscope. This exposes them to pollutants and changes in thesurrounding environment and may jeopardize the integrity of the experiment.This thesis presents the development of a cost-effective, miniaturized microscope designedfor imaging of cell cultures directly within incubators. By integrating simple, inexpensiveglass lenses and 3D-printed components and focusing on the ESP32-CAM module for digitalimaging, the project explores various optical setups to optimize image quality while minimizingdisruption to cell environments.Central to the research was the identification and testing of diverse optical configurations todetermine the most effective arrangement for both brightfield and fluorescence microscopy.The design features a baseplate for stability, a filter plate for fluorescence imaging, and afocus adjustment mechanism using magnets. Iterative enhancements led to a side illuminationtechnique using an economical LED, removing the need for a beamsplitter and simplifying theoptical path.The final microscope demonstrated successful brightfield imaging and weak fluorescenceimaging of Madin-Darby Canine Kidney (MDCK) II cell cultures marked with Green FluorescentProtein (GFP), using a magnification ratio of 2.5:1 through an infinity-corrected optical system.The findings illustrate the potential of developing an economical, functional microscope thatcan be readily replicated and scaled for use in cell culture technology. / En central del av biologisk forsking är användningen av cellkulturer, vilket tillåter forskare attutforska beeendet och funktionerna av celler i kontrollerade miljöer. Konventionell bevakning avcellkulturer kräver ofta att de tas ut från deras inkubatorer för att observeras under ett mikroskop.Detta kan utsätta dem för föroreningar och förändringar i omgivningen vilket kan äventyra helaexperimentet.Det här examensarbetet beskriver utvecklingen av ett kostnadseffektivt, miniatyriserat mikroskopanpassat för att avbilda cellkulturer inuti inkubatorer. Genom att integrera enkla, billigaglaslinser och 3D-printade komponenter, samt ESP32-CAM modulen för bildtagning, utforskardetta arbete olika optiska system för att optimera bildkvalitet och minimera störningar i cellernasmiljö.En väsentlig del av forskningen involverade identifiering och testning av olika optiska konfigurationerför att bestämma det mest effektiva arrangemanget för både brightfield- och fluorescensmikroskopi.Designen inkluderar en basplatta för stabilitet, en filterplatta för fluorescensavbildning ochen fokusjustering som utnyttjar magneter. Iterativa förbättringar ledde till utvecklingen av enbelysningsteknik med en billig LED, vilket tog bort behovet av en stråldelare och förenkladeden optiska banan.Det slutgiltiga mikroskopet uppvisade framgångsrik avbildning med brightfield och begränsadavbildning med fluorescens av MDCK II cellkulturer märkta med GFP. En förstoringsfaktor2.5:1 användes genom ett oändlighetskorrigerat optiskt system. Resultaten demonstrerar potentialenav att utveckla ett ekonomiskt och funktionellt mikroskop som kan replikeras för användninginom cellkulturforskning.

Molecules adsorbed on two-dimensional (2D) materials can show interesting physical and chemical properties. This thesis presents scanning probe microscopy (SPM) investigation of emerging 2D materials, molecular nanostructures on 2D substrates at the nanometer scale, and biophysical processes on the biological membrane. Two main techniques of nano-probing are used: scanning tunneling microscopy (STM) and atomic force microscopy (AFM). The study particularly emphasizes on self-assembled molecules on flat 2D materials and quasi-1D wrinkles. First, we report the preparation of novel 1D C60 nanostructures on rippled graphene. Through careful control of the subtle balance between the linear periodic potential of rippled graphene and the C60 surface mobility, we demonstrate that C60 molecules can be arranged into a 1D C60 chain structure of two to three molecules in width. At a higher annealing temperature, the 1D chain structure transitions to a more closely packed, quasi-1D hexagonal stripe structure. The experimental realization of 1D C60 structures on graphene is, to our knowledge, the first in the field. It could pave the way for fabricating new C60/graphene hybrid structures for future applications in electronics, spintronic and quantum information. Second, we report a study on nano-morphology of potential operative donors (e.g., C60) and acceptors (e.g., perylenetetracarboxylic dianhydride, aka. PTCDA) on wrinkled graphene supported by copper foils. We realize sub-monolayer C60 and PTCDA on quasi-1D and quasi-2D real periodic wrinkled graphene, by carefully controlling the deposition parameters of both molecules. Our successful realization of acceptor-donor binary nanostructures on wrinkled graphene could have important implications in future development of organic solar cells. Third, we report an STM and spectroscopy study on atomically thin transition-metal dichalcogenides (TMDCs) material. TMDCs are emerging 2D materials recently due to their intriguing physical properties and potential applications. In particular, our study focuses on molybdenum disulfide (MoS2) mono- to few-layers and pyramid nanostructures synthesized through chemical vapor deposition. On the few-layered MoS2 nanoplatelets grown on gallium nitride (GaN) and pyramid nanostructures on highly oriented pyrolytic graphite (HOPG), we observe an intriguing curved region near the edge terminals. The measured band gap in these curved regions is consistent with the direct band gap in MoS2 monolayers. The curved features near the edge terminals and the associated electronic properties may contribute to understanding catalytic behaviors of MoS2 nanostructures and have potential applications in future electronic devices and catalysts based on MoS2 nanostructures. Finally, we report a liquid-cell AFM study on the endosomal protein sorting process on the biological lipid membrane. The sorting mechanism relies on complex forming between Tom1 and the cargo sorting protein, Toll interacting protein (Tollip). The induced conformational change in Tollip triggers its dissociation from the lipid membrane and commitment to cargo trafficking. This collaborative study aims at characterizing the dynamic interaction between Tollip and the lipid membrane. To study this process we develop the liquid mode of AFM. We successfully demonstrate that Tollip is localized to the lipid membrane via association with PtdIns3P (PI(3)P), a major phospholipid in the cell membrane involved in protein trafficking. / Ph. D. / Two-dimensional (2D) materials are layered materials with thickness of single atom or few atoms. The ultimate thickness leads to novel properties that are useful for a wide range of applications in photovoltaics, electronics and quantum information. In order to explore these properties at the nanometer scale, we used scanning probe techniques, i.e., scanning tunneling microscopy (STM) and atomic force microscopy (AFM), to perform comprehensive investigations on these emerging materials. 2D materials, such as graphene and atomically thin transition-metal dichalcogenides (TMDCs), are promising candidates for building economic, safe and mechanically flexible solar cells with desirable optical and electronic properties, e.g. tunable sunlight absorption. The first part of the thesis focuses on graphene, a single-atom-thick carbon sheet. We deposited key components in organic solar cells, such as perylenetetracarboxylic dianhydride (PTCDA) and C₆₀ molecules, on graphene. On these materials we observed various novel nanostructures, like quasi-1D C₆₀ nanochains. The second part of the thesis focuses on mono- to few-layered MoS₂, which can be used as an active layer in high-efficiency solar cells. Our study has important implications in improving efficiency of organic solar cells in the future. In the final part of the thesis, we extended our subject to the biological lipid membrane, a 2D material critical in biology, and biophysical processes occurring on the membrane. Using a liquid-cell AFM, we investigated the endosomal protein sorting process on the biological membranes. Our study contributes to understanding of the interactions between the adaptor proteins and cell membranes in the protein sorting process that guides proteins to their proper destinations.

In life science practices, cell culturing is one of the most common procedures in biological experiments involving the use of microscopes. Cell cultures are typically viewed with a microscope to monitor changes after adjusting the environment for the cells or to intermittently supervise for cellular behavior. However, it is currently not possible to continuously monitor cellular behavior due to the absence of solutions for wireless control of a microscope to image cell cultures while maintaining the environment conducive to cell growth. This thesis provides a miniaturized, cost-effective and versatile microscope down-scaled enough to operate inside incubators. This versatility encompasses the ability to switch between bright-field mode and fluorescence mode, capture images in sequences, save said sequential images to a micro SD-card as well as enabling downloading of individual images to a local file manager. All these features are done through a user interface set up by either a local WiFi or access point from the microscope, making the operation possible through wireless connection. In addition, this thesis is a continuation of a previous thesis that was completed in the spring of 2023. The purpose of this thesis is to improve on the result of the main research question of the previous work. / Inom livsvetenskapliga metoder är cellodling en av de vanligaste procedurerna i biologiska experiment som involverar användning av mikroskop. Cellkulturer ses vanligtvis med ett mikroskop för att bevaka förändringar efter att ha ändrat miljön för cellerna eller för att intermittent övervaka cellulärt beteende. Men det är för närvarande inte möjligt att kontinuerligt övervaka cellulärt beteende på grund av avsaknaden av lösningar för trådlös kontroll av ett mikroskop för att avbilda cellkulturer samtidigt som miljön som främjar celltillväxt bibehålls. Detta examensarbete tillhandahåller ett miniatyriserat, kostnadseffektivt och mångfasetterat mikroskop, som är tillräckligt nedskalad för att fungera i inkubatorer. Denna mångsidighet omfattar möjligheten att växla mellan ljusfältsläge och fluorescensläge, ta bilder i sekvenser, spara nämnda sekventiella bilder på ett micro SD-kort samt ladda ner enskilda bilder till en lokal filhanterare. Alla dessa funktioner görs via ett användargränssnitt som ställs in av antingen ett lokalt WiFi eller accesspunkt från mikroskopet, vilket gör operationen möjlig via trådlös anslutning. Dessutom är detta examensarbete en fortsättning på ett tidigare examensarbete som avslutades våren 2023. Syftet med detta examensarbete är att förbättra resultatet på den huvudsakliga forskningsfrågan i det tidigare arbetet.

As sessile organisms, plants manage stress through complex signaling networks involving phytohormones such as phytosulfokine (PSK). PSK, a disulfated pentapeptide, regulates plant growth, development, and stress responses by interacting with specific PSK receptors (PSKRs). In this study, we explored the trafficking dynamics of PSK, its post-application fate, and the synthesis of an analog. We administered both native PSK and a fluorescent version tagged with TAMRA (5(6)-carboxytetramethylrhodamine) to various Arabidopsis thaliana genotypes, including wild type, a PSKR-deficient mutant, and a strain overexpressing PSKR1 tagged with green fluorescent protein (GFP) over the wild-type background. Fiber-optic fluorescence microscopy revealed that receptor presence influences PSK’s internal movement. Additionally, we extracted TAMRA–PSK from treated plants and recovered it using solid-phase extraction to assess its stability post-application. HPLC analysis suggested that TAMRA–PSK is substantially unchanged in the plant matrix. Furthermore, a PSK analog was partially synthesized via solid-phase peptide synthesis for future studies.

This thesis describes the physical nature of corrections of an electron microscope and mathematical methods of image processing required for their complete automation. The corrections include different types of focusing, astigmatism correction, electron beam centring, and image stabilisation. The mathematical methods described in this thesis include various methods of measuring focus and astigmatism, with and without using the Fourier transform, edge detection, histogram operations, and image registration, i.e. detection of spatial transformations in images. This thesis includes detailed descriptions of the mathematical methods, their evaluation using an “offline” application, descriptions of the algorithms of their implementation into an actual electron microscope and results of their testing on the actual electron microscope, in the form of a video footage grabbed from its control computer’s screen.