Pablo Rivera Fuentes


Last Name

Rivera Fuentes

First Name

Pablo

ORCID

0000-0001-8558-2828

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2018 - 201982020 - 20259
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Publications 1 - 10 of 17
  • Photoregulated fluxional fluorophores for live-cell super-resolution microscopy with no apparent photobleaching
    Item type: Journal Article
    Halabi, Elias A.; Pinotsi, Dorothea; Rivera Fuentes, Pablo (2019)
    Nature Communications
    Photoswitchable molecules have multiple applications in the physical and life sciences because their properties can be modulated with light. Fluxional molecules, which undergo rapid degenerate rearrangements in the electronic ground state, also exhibit switching behavior. The stochastic nature of fluxional switching, however, has hampered its application in the development of functional molecules and materials. Here we combine photoswitching and fluxionality to develop a fluorophore that enables very long (>30 min) time-lapse single-molecule localization microscopy in living cells with minimal phototoxicity and no apparent photobleaching. These long time-lapse experiments allow us to track intracellular organelles with unprecedented spatiotemporal resolution, revealing new information of the three-dimensional compartmentalization of synaptic vesicle trafficking in live human neurons.
  • Revitalizing antibiotic discovery and development through in vitro modelling of in-patient conditions
    Item type: Other Journal Item
    Sollier, Julie; Basler, Marek; Broz, Petr; et al. (2024)
    Nature Microbiology
    In vitro models mimicking in-patient conditions have the potential to yield exciting opportunities for antibiotic research and revitalize future antibiotic discovery and development.
  • Synthesis of Asparagusic Acid Modified Lysine and its Application in Solid-Phase Synthesis of Peptides with Enhanced Cellular Uptake
    Item type: Journal Article
    Tirla, Alina; Hansen, Moritz; Rivera Fuentes, Pablo (2018)
    Synlett
  • 12th Young Faculty Meeting, 28th May 2019
    Item type: Other Journal Item
    Šolomek, Tomáš; Rivera Fuentes, Pablo; Merz, Leo (2019)
    Chimia
  • Rapid Characterization and Quantification of Extracellular Vesicles by Fluorescence‐Based Microfluidic Diffusion Sizing
    Item type: Journal Article
    Paganini, Carolina; Hettich, Britta; Kopp, Marie R.G.; et al. (2022)
    Advanced Healthcare Materials
    Extracellular vesicles (EVs) are emerging as promising diagnostic and therapeutic tools for a variety of diseases. The characterization of EVs requires a series of orthogonal techniques that are overall time- and material-consuming. Here, a microfluidic device is presented that exploits the combination of diffusion sizing and multiwavelength fluorescence detection to simultaneously provide information on EV size, concentration, and composition. The latter is achieved with the nonspecific staining of lipids and proteins combined with the specific staining of EV markers such as EV-associated tetraspanins via antibodies. The device can be operated as a single-step immunoassay thanks to the integrated separation and quantification of free and EV-bound fluorophores. This microfluidic technique is capable of detecting and quantifying components associated to EV subtypes and impurities and thus to measure EV purity in a time scale of minutes, requiring less than 5 µL of sample and minimal sample handling before the analysis. Moreover, the analysis is performed directly in solution without immobilization steps. Therefore, this method can accelerate screening of EV samples and aid the evaluation of sample reproducibility, representing an important complementary tool to the current array of biophysical methods for EV characterization, particularly valuable for instance for bioprocess development.
  • The Bioorthogonal Isonitrile-Chlorooxime Ligation
    Item type: Journal Article
    Schäfer, Rebecca J.B.; Monaco, Mattia R.; Li, Mao; et al. (2019)
    Journal of the American Chemical Society
  • Characterization of the glutathione redox state in the Golgi apparatus
    Item type: Journal Article
    Miró-Vinyals, Carla; Emmert, Sarah; Grammbitter, Gina; et al. (2025)
    Redox Biology
    Redox homeostasis is crucial for cell function, and, in eukaryotic cells, studying it in a compartmentalized way is essential due to the redox variations between different organelles. The redox state of organelles is largely determined by the redox potential of glutathione, EGSH, and the concentration of its reduced and oxidized species, [GS]. The Golgi apparatus is an essential component of the secretory pathway, yet little is known about the concentration or redox state of GSH in this organelle. Here, we characterized the redox state of GSH in the Golgi apparatus using a combination of microscopy and proteomics methods. Our results prove that the Golgi apparatus is a highly oxidizing organelle with a strikingly low GSH concentration (EGSH = – 157 mV, 1–5 mM). These results fill an important gap in our knowledge of redox homeostasis in subcellular organelles. Moreover, the new Golgi-targeted GSH sensors allow us to observe dynamic changes in the GSH redox state in the organelle and pave the way for further characterization of the Golgi redox state under various physiological and pathological conditions.
  • A Molecular Logic Gate Enables Single-Molecule Imaging and Tracking of Lipids in Intracellular Domains
    Item type: Journal Article
    Eördögh, Ádám; Paganini, Carolina; Pinotsi, Dorothea; et al. (2020)
    ACS Chemical Biology
    Photoactivatable dyes enable single-molecule imaging and tracking in biology. Despite progress in the development of new fluorophores and labeling strategies, many intracellular compartments remain difficult to image beyond the limit of diffraction in living cells. For example, lipid domains, e.g., membranes and droplets, remain difficult to image with nanometric resolution. To visualize these challenging subcellular targets, it is necessary to develop new fluorescent molecular devices beyond simple on/off switches. Here, we report a fluorogenic molecular logic gate that can be used to image single molecules associated with lipid domains, most notably droplets, with excellent specificity. This probe requires the subsequent action of light, a lipophilic environment, and a competent nucleophile to produce a fluorescent product. The combination of these inputs results in a probe that can be used to image the boundary of lipid droplets in three dimensions with resolution beyond the limit of diffraction. Moreover, this probe enables single-molecule tracking of lipid trafficking between droplets and the endoplasmic reticulum.
  • Dual-Activatable Cell Tracker for Controlled and Prolonged Single-Cell Labeling
    Item type: Journal Article
    Halabi, Elias A.; Arasa, Jorge; Püntener, Salome; et al. (2020)
    ACS Chemical Biology
    Cell trackers are fluorescent chemical tools that facilitate imaging and tracking cells within live organisms. Despite their versatility, these dyes lack specificity, tend to leak outside of the cell, and stain neighboring cells. Here, we report a dual-activatable cell tracker for increased spatial and temporal staining control, especially for single-cell tracking. This probe overcomes the typical problems of current cell trackers: off-target staining, high background signal, and leakage from the intracellular medium. Staining with this dye is not cytotoxic, and it can be used in sensitive primary cells. Moreover, this dye is resistant to harsh fixation and permeabilization conditions and allows for multiwavelength studies with confocal microscopy and fluorescence-activated cell sorting. Using this cell tracker, we performed in vivo homing experiments in mice with primary splenocytes and tracked a single cell in a heterogeneous, multicellular culture environment for over 20 h. These experiments, in addition to comparative proliferation studies with other cell trackers, demonstrated that the signal from this dye is retained in cells for over 72 h after photoactivation. We envision that this type of probes will facilitate the analysis of single-cell behavior and migration in cell culture and in vivo experiments.
  • Disruption of mitochondrial redox homeostasis by enzymatic activation of a trialkylphosphine probe
    Item type: Journal Article
    Nguyen, Jade; Tirla, Alina; Rivera Fuentes, Pablo (2021)
    Organic & biomolecular chemistry
    Redox homeostasis is essential for cell function and its disruption is associated with multiple pathologies. Redox balance is largely regulated by the relative concentrations of reduced and oxidized glutathione. In eukaryotic cells, this ratio is different in each cell compartment, and disruption of the mitochondrial redox balance has been specifically linked to metabolic diseases. Here, we report a probe that is selectively activated by endogenous nitroreductases, and releases tributylphosphine to trigger redox stress in mitochondria. Mechanistic studies revealed that, counterintuitively, release of a reducing agent in mitochondria rapidly induced oxidative stress through accumulation of superoxide. This response is mediated by glutathione, suggesting a link between reductive and oxidative stress. Furthermore, mitochondrial redox stress activates a cellular response orchestrated by transcription factor ATF4, which upregulates genes involved in glutathione catabolism.
Publications 1 - 10 of 17