Development of new applications of inductively coupled plasma mass spectrometry (ICP-MS) hyphenated with different sample introduction systems

225 p. : il. Texto en español con conclusiones en inglés

La cerámica de la Segunda Edad del Hierro en el País Vasco: Estudio tecnológico, funcional y social

670 p. Capítulos de introducción, metodología, discusión y conclusiones en castellano e inglés.

Advantages and Versatility of Fluorescence-Based Methodology to Characterize the Functionality of LDLR and Class Mutation Assignment

Familial hypercholesterolemia (FH) is a common autosomal codominant disease with a frequency of 1:500 individuals in its heterozygous form. The genetic basis of FH is most commonly mutations within the LDLR gene. Assessing the pathogenicity of LDLR variants is particularly important to give a patient a definitive diagnosis of FH. Current studies of LDLR activity ex vivo are based on the analysis of I-125-labeled lipoproteins (reference method) or fluorescent-labelled LDL. The main purpose of this study was to compare the effectiveness of these two methods to assess LDLR functionality in order to validate a functional assay to analyse LDLR mutations. LDLR activity of different variants has been studied by flow cytometry using FITC-labelled LDL and compared with studies performed previously with I-125-labeled lipoproteins. Flow cytometry results are in full agreement with the data obtained by the I-125 methodology. Additionally confocal microscopy allowed the assignment of different class mutation to the variants assayed. Use of fluorescence yielded similar results than I-125-labeled lipoproteins concerning LDLR activity determination, and also allows class mutation classification. The use of FITC-labelled LDL is easier in handling and disposal, cheaper than radioactivity and can be routinely performed by any group doing LDLR functional validations.

Measurement of polycyclic aromatic hydrocarbons in ambient air by fluorescence spectroscopic techniques

250 p. + anexos

Mass spectrometry coupled to imaging techniques: The better the view the greater the challenge

These are definitively exciting times for membrane lipid researchers. Once considered just as the cell membrane building blocks, the important role these lipids play is steadily being acknowledged. The improvement occurred in mass spectrometry techniques (MS) allows the establishment of the precise lipid composition of biological extracts. However, to fully understand the biological function of each individual lipid species, we need to know its spatial distribution and dynamics. In the past 10 years, the field has experienced a profound revolution thanks to the development of MS-based techniques allowing lipid imaging (MSI). Images reveal and verify what many lipid researchers had already shown by different means, but none as convincing as an image: each cell type presents a specific lipid composition, which is highly sensitive to its physiological and pathological state. While these techniques will help to place membrane lipids in the position they deserve, they also open the black box containing all the unknown regulatory mechanisms accounting for such tailored lipid composition. Thus, these results urges to different disciplines to redefine their paradigm of study by including the complexity revealed by the MSI techniques.

Estudio de la interacción entre los canales Kv7.2 y la calmodulina utilizando la técnica Bimolecular Fluorescence Complementation (BiFC)

Empleo de la técnica BiFC para estudiar la interacción entre los canales Kv7.2 y la CaM. Para ello se realizan clonajes, se co-transfectan distintas combinaciones de éstos en células de mamífero y se analiza y cuantifica la intensidad de fluorescencia obtenida mediante microscopía confocal.

Fluorescence cross-correlation spectroscopy to study hemodynamics

[EN] In the recent years a series of optical correlation techniques have been developed in order to be able to measure flow velocity with high spatial resolution while being non-invasive in order to be employed in-vivo on biological organisms. The technique employed in my thesis work, scanning laser image correlation (SLIC), is a powerful approach for the detection of flow motions because it overcomes some limitations of the classical spectroscopy techniques. SLIC method consists in repeated laser scans over a linear pattern and on the cross correlation of the signal emitted by the excited fluorophores in different positions along the scan line. Therefore, the resulting measurements for flow velocity are really accurate.

Effect of surface plasmon resonance in TiO2/Au thin films on the fluorescence of self-assembled CdTe QDs structure

23rd Congress of the International Comission for Optics (ICO 23)