Platforms

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 (Horiba T64000). 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.
  • Raman Microspectroscopy System (Thermo Scientific DXR2). The Thermo Scientific™ DXR2™ Raman microscope delivers very high spatial resolution, together with the reliability, performance and reproducibility required in demanding analytical environments. Designed for versatility, the DXR2 Raman microscope makes it as easy as possible to get high quality results. The DXR Raman microscope is equipped with: a) class I laser-safe design for use in non-restricted environments; b) two excitation wavelengths (633nm 785nm) for optimal results with demanding biological samples.; c) patented auto-alignment system for highest optimal performance; d) rapid, automated, multi-point calibration system for confidence in sample identification; e) 1 µm x, y spatial resolution and 2 µm depth resolution; f) regulator Laser Power to assure reproducible laser power at sample; g) 10X and 50X optical objectives. Lasers are pre-aligned and user exchangeable, no tools required. All lasers are depolarized which minimizes the sample orientation dependence in measurements.
  • 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 time-resolved 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.
  • Quanterix SR-X: ultra-sensitive biomarker detection system. The system is powered by Simoa® bead technology, and offers researchers access to biomarker detection capabilities in a compact and affordable system with sensitivity into the femtogram (fg/ml) range, representing up to 1,000 times higher sensitivity than traditional enzyme-linked immunosorbent assay (ELISA). The SR-X is designed for multiplex detection of up to six analytes per well, with low volume requirements to increase productivity and throughput, while conserving your precious samples. An extensive menu of Simoa assay kits are available to measure critical biomarkers with substantially higher sensitivity than standard immunoassay methods, enabling detection of both normal and acute levels with high precision across a range of sample types.
  • The NanoSight NS300 uses the technology of Nanoparticle Tracking Analysis (NTA). This technology utilizes the properties of both light scattering and Brownian motion in order to obtain the size distribution and concentration measurement of particles in liquid suspension (20-2000nm). The technique is used in combination with a microscope that allows you to visualize the movement of small particles under Brownian motion. Dedicated software for tracking particle movements and evaluating their hydrodynamic diameters using the Stokes-Einstein equation. The NTA technology allows high resolution dimensional distribution measurements, the relative scattering intensity of each particle, concentration and aggregation measurements and, thanks to the fluorescence mode, to obtain specific data for the marked particles (Laser equipment: blue 488nm and green 532nm).
  • MicroCal PEAQ-ITC The MicroCal PEAQ-ITC instrument is a highly sensitive semi-automatic isothermal titration microcalorimetry system with a micro-volume cell. It is characterized by the automatic loading of the titration syringe and by the integrated automatic washing module. The system is dedicated to the study of biomolecular interactions, and it does not require the labelling or immobilization of the interacting molecules and has no molecular mass limitations. The instrument provides a direct measure of the affinity constant from millimolar to nanomolar values ​​(from 10-2 to 10-9 M) and it can also be used to measure enzymatic kinetics, interactions with nanomaterials and other interactions between components in solution.

  • Spectrofluorometer JASCO FP-8500    The FP-8500 is a high performance spectrofluorometer with high sensitivity and spectral accuracy (1nm). With a water Raman S/N ratio of 8,500:1 (RMS), it allows fast measurements of samples with low level fluorescence.The high-speed scanning enables fast measurements of 3D spectra and phosphorescence samples.It operates in a 200 to 850nm range.


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