Spectroscopy, diffractometry and molecular interaction study platform

It fulfills the requirements of characterizing organic and inorganic molecules, macromolecules and their interactions.

  • Nuclear Magnetic Resonance (NMR) Spectrometer. The instrumentation consists of a high resolution NMR spectrometer with a triple resonance cryogenic probe. The magnet is a Bruker Avance III 600 Ultrashield plus spectrometer operating at 600 MHz (14.1 Tesla) and it is equipped with a TCI Cryoprobe in triple resonance (enables enhanced sensitivity and optimized for 1H and 13C observation) and with an Automatic Sample Changer B-ACS 60 (enables a high sample throughput in the daily routine or overnight automation). The NMR spectroscopy has a wide range of applications including the evaluation and the analysis of small organic molecules, the characterization of biological macromolecules at atomic level (proteins, nucleic acids), protein-ligand and protein-protein interaction studies, metabolomics based studies of biological fluids (urine, serum, saliva, CSF), nanobiotechnology, quality control and food analysis.
  • Raman Microspectroscopy System. The apparatus consists of an Ar/Kr Gas Laser (Stabilite RM2018, Physics spectra) with a cooling system water supply, having a power of 3W, capable of providing about ten excitation lines in the visible region (450-670nm). The Laser is coupled with triple monochromator based Raman spectrometer (T-64000, Horiba-Jobin Yvon) equipped with an Olympus microscope (BX-41) with three objectives (10x, 50x and 100x) and with a liquid nitrogen-cooled CCD detector (256 × 1024 pixel). Excellent stray light rejection can be achieved when the double subtractive filtering mode is used. This configuration allows spectra of low frequency bands very close to the laser line (typically down to 5 cm-1) over a broad wavelength range. This instrument is used for studies of vibrational dynamics of single crystals, but can also be used to characterize micro and nano-composite structured systems, in the spectral range of mid and far-infrared with lateral spatial resolution of 1 micron and with spectral resolution of less than 0.5 cm-1/ pixel.
  • X-ray powder diffractometer. The instrument is equipped with a copper anode x-ray tube (Kα, λ = 1.5418 Å) and a Peltier solid-stage detector that allows to cool down the Si(Li) crystal down to a temperature close to -100°C and therefore, the internal noise of this detector is extremely low. The instrument can be used to obtain detailed structural and chemical information about the crystallography of materials (organic and inorganic) in the form of polycrystalline powders or thin films. The instrument has important applications in the fields of materials and pharmaceutical science. In the former case, the crystalline structure/property relationship of the material and the problem of polymorphic forms of drugs in the latter, are extremely important topics in the chemical and biochemical fields.
  • Modular fluorescence spectrometer. The instrument allows fluorescence analysis, in continuous wave and timeresolved regimes, of samples both in solution and in the solid state. It consists of a modular spectrofluorometer (Nanolog/Fluorolog-3-2iHR320, Horiba-Jobin Yvon) equipped with Xenon lamp (450W, ozone free) completed of current-stabilized power supplies. It can also be used for life-time measurements and for quantum yield measurements.
  • Zetasizer Nano ZS (Malvern). The system is used for the measurement of the size (Dynamic Light Scattering), zeta potential and electrophoretic mobility (Laser Doppler Microelectrophoresis) of colloids and nanoparticles, and molecular weight (Static Light Scattering) of proteins and polymer solutions.
  • Circular Dichroism Spectrophotometer (Jasco J-1500). The system allows the measurement of CD spectra in the UV-Vis region (163-950nm) and with a spectral bandwidth of 01-16 nm. The instrument includes a Peltier temperature control system which enables to measure the CD spectrum with change in the sample temperature from -30 to 130 °C. The double polarizing prism monochromators of the spectrophotometer result in low stray light enabling the instruments to obtain high-quality CD data even under conditions with high absorbance. The addition of a dedicated monochromator and detector provides simultaneous acquisition of fluorescence and CD measurements. The instrument is equipped with the Spectra Manager II software suite for designing experimental methods, acquiring data and processing results. Analysis programs designed for CD data include Multivariate SSE, Curve fitting, Denaturated protein analysis, Protein ligand analysis e Protein SSE analysis. CD Spectrometer system allows the analysis of the structural properties of biomolecules and optically active compounds, and it can be used to estimate the secondary structure content (α-helix, β-structure, random coil), to monitor conformational changes of proteins upon their interaction with ligand molecules or for following the unfolding and folding of proteins as a function of temperature.
  • Dual-channel automatic SPR Pioneer SensiQ (ICx Nomadics). The surface plasmon resonance (SPR) is a suitable technique to characterize a wide variety of biomolecular interactions. SPR provides non-invasive, label-free observation of binding interactions between an injected analyte and an immobilized biomolecule in real time (protein-ligand or protein-protein interactions). The SensiQ system can be used to confirm the binding events and to determine the affinity, stoichiometry and kinetic parameters of association and dissociation Moreover, using a calibration curve, the active analyte concentration can be calculated from the SPR data.
  • Nano Isothermal Titration Calorimeter (Nano-ITC, TA Instruments). The Isothermal Titration Calorimetry (ITC) is a thermodynamic tool for label-free analysis of macromolecular interactions in solution. The technique is suitable for analyzing protein-protein, protein-DNA interactions as well as interactions between macromolecules and small ligands. By measuring the amount of heat released or taken up during each reaction in a titration experiment, the equilibrium binding affinity and the stoichiometry are derived. In addition, ITC is the only technique able to directly measure the thermodynamic parameters of binding such as enthalpy, entropy. The Nano-ITC instrument has the advantage to use very small volumes of often precious biological samples (cell volume at 190uL), to minimize sample consumption and at the same time provides high sensitivity levels required for characterizing the fundamental driving forces of biomolecular binding reactions.
  • Differential scanning calorimeter (Nano-DSC, TA Instruments). The differential scanning calorimetry (DSC) is a technique used to determine the thermal stability and heat capacity of proteins and other bio-molecules (e.g. DNA) in its native form (thermal transition or melting temperature). DSC measures the difference in the amount of heat absorbed or released by sample and reference as they are heated or cooled at constant pressure. The Nano-DSC is specifically designed to determine the thermal stability and heat capacity of proteins in dilute solution, with the versatility and precision to perform molecular stability screening and to characterize both the specific or nonspecific ligand binding.
  • Stopped-flow system. Stopped-flow spectrophotometry is an analytical technique used in the study of fast chemical and enzymatic kinetics or fast reactions on the millisecond timescale. The stopped-flow instrument incorporates several modules connected with each other. A rapid mixing device (SFM-300 BioLogic), controlled by the microprocessor unit (MPS-60 BioLogic) forces two or three solutions (contained in syringes) together through a high efficiency mixing chamber into an absorption cell. The system uses a xenon lamp for UV and visible wavelengths and a diode-array spectrometer (J&M Analytik), which enables quick and precise measuring with a low level of scattered light. The configuration of the system can be converted quickly to perform quench-flow experiments.