Development of BIOnanotechnologies based on Extracellular Vesicles for early diagnosis, prognosis and therapy of atherosclerotic disease (BIOVEA)
Various biomarkers have been proposed for the diagnosis and prognosis of acute coronary syndrome, however none for stable coronary disease. Identification of biomarkers that augment established clinical indicators, may allow earlier diagnosis of coronary disease, as well as improved recognition of the risk of complications and cardiovascular death. Additionally, this would pave the way for biomarkers, such as extracellular vesicles, to be employed as precise therapeutic agents in these diseases.
The Ultrastructural Pathology and Bioimaging Lab has outstanding research equipment, both newly acquired as well as purchased and upgraded over the years. These cutting edge tools are essential for improved diagnostics in various human diseases, and also for gaining new insights into cellular functions, at the molecular level.
The 100 kV FEI Morgagni Transmission Electron Microscope (TEM), equipped with an Olympus MegaView III CCD, is our main diagnostic tool, allowing fast and qualitative evaluation of plastic embedded specimens. It is also occasionally used for demonstrations and teaching.
The FEI T12 is a versatile 120 kV TEM, employed for both routine diagnostic, as well as for more demanding applications, such as tomography of chemically fixed specimens. At its heart lies a Lab6 filament and image acquisition is done by an Olympus MegaView G2 CCD. Recently, this microscope was fitted with a Tecnai iCorr module for correlative light and electron microscopy (CLEM), which allows merging of fluorescence and transmission electron microscopy images.
The 200 kV FEI Talos F200C TEM enables our researchers to visualize cellular structures and isolated macromolecular assemblies in remarkable detail, in 2 and 3-dimensions. This TEM is equipped to handle metal coated and chemically fixed specimens, but also cryogenically frozen samples, a technique which preserves the native, hydrated structure. It is currently the only cryo-TEM in Romania dedicated to life-sciences applications. This system was acquired through POSCCE 2.2.1 Grant 633/2014 (ARMED).
Our preparation rooms have tools for (almost) every TEM life-science application and user need. They house two cryo-plungers (Leica EM GP and FEI Vitrobot Mark IV), a high pressure freezer (Leica EM PACT2), room temperature and cryo ultramicrotomes (RMC MT XL and two Leica EM UC7), a freeze substitution system (Leica EM AFS2), a coating unit (Quorum Technologies Q150 ES) and a number of other ancillaries. Whether attempting plastic-embedding of tissue specimens or cryo-TEM studies of cultured cells grown on grids or single particles (e.g. proteins or viruses), this equipment can deliver reproducible samples. Most were acquired through POSCCE 2.2.1 Grant 633/2014 (ARMED).
The Leica TCS SP8 STED3X Super-Resolution Microscope (currently the only one of its kind in Romania) is the latest iteration of a Nobel Prize winning technology, one that literally sheds new light on the dynamics of fluorescently-tagged molecules in living cells. It uses Stimulated Emission Depletion (STED), in which a laser selectively activates and deactivates fluorophores to obtain nanoscale resolution, thereby overcoming the limits of optical microscopy. This system was acquired through POSCCE 2.2.1 Grant 633/2014 (ARMED).
Our cell biology research focuses not only on bioptic tissue samples obtained from patients, but extends to live cell studies. The tissue culture facility offers the necessary logistics for cell culture, transfections and in the future, the use of viral vector systems. The tissue culture facility houses a biology safety cabinet, 2 incubators, Invitrogen EVOS microscopes, a number of centrifuges (including a Thermo Scientific Sorvall WX+ ultracentrifuge) and other ancillary items. Equipment was acquired through POSCCE 2.2.1 Grant 633/2014 (ARMED).
