Instrumentation

Optical Microscopy Equipment

Confocal fluorescence microscope, Zeiss LSM 880 Airyscan and MaiTai DeepSee.
The microscope is equipped with three Ar laser excitation sources (458, 488 and 514 nm), DPSS (561 and 590 nm) and HeNe (633 nm). It is coupled to a MaiTai DeepSee multiphoton laser, with variable excitation range (690 to 1040 nm). It has two PMT photomultiplier type detectors, one standard and one with high sensitivity in the infrared range, as well as a GaAsP detector for detection in the visible spectrum. All three spectral detectors can be used simultaneously in confocal image acquisition. The Airyscan module is an array of detectors that allows measurements with high lateral resolution of 150 nm and axial of 400 nm, on the 488 channels. It also has a BIG-2 detector module, composed of two GaAsP detectors for high sensitivity multiphoton measurements. Allows for fluorescence correlation and cross-correlation spectroscopy measurements.

Confocal fluorescence microscope ZEISS LSM 900 Light Scanning Microscope (Zeiss 900) it´s a fully automatized inverted fluorescence confocal microscope. Allows the acquisition of high-resolution images of 3 fluorescence channels in simultaneous mode. It´s fitter with AI Sample finder for faster image localization and experiment implementation routines. Allows Z-stack, time series and tile acquisition, FRAP and FRET experiments and different routines for automatization. Microscope can be fitted with 2 inserts, for multiwell dishes (universal frame) and for single petri/glasslide preparations. System runs on full licenced ZEN blue software including experimental designer, LSMPlus, Tile & Position module, linear unmixing, colocalization and 3D Viewer and processing toolkit.

Epifluorescence microscope, Zeiss Axio Cell Observer. It has a lighting system consisting of four Colibri LEDs (365, 470, 530 and 590 nm) and a Xe lamp. It is equipped with two AxioCam cameras: MRm3 and 305color. The microscope is equipped with an incubation cabinet for experiments with temperature, CO2 and humidity control, which allows long-term in vitro experiments. The system is controlled with the software Zen Blue.

Raman confocal microscope, Renishaw inVia Reflex. Allows the analysis of solids or substrates immersed in water, as well as liquid or suspended samples. The Leica microscope integrated in the system is equipped with air and water immersion objectives, a motorized scanning sample platform and two video cameras. It is coupled with three laser excitation sources at 532, 633 and 785 nm (35, 17 and 120 mW). The spectrometer features front-illuminated (FI) CCD and back-illuminated deep-depletion (BI-DD) CCD operating with diffraction gratings of 2400, 1800, 1200 and 600 lines/mm providing ultra-high spectral resolution < 1 cm-1 for 532 nm, and high (1 cm-1) or low spectral resolution options for all wavelengths. In addition, Streamline optics, accessory for macroscopic samples, temperature and humidity cell are available. Polarisation can be varied by 2deg steps for 633 and 785 nm, orthogonal and circular filters are installed for all wavelengths. The system is controlled with the software Wire 5.6.

Raman confocal microscope, WITec alpha300 R. Allows the analysis of solids, liquids or substrates immersed in water. The integrated Zeiss microscope is equipped with different, air and water immersion objectives, a CMOS video camera and a topographic surface sensor that allows measuring on rough substrates while maintaining focus. It has four laser excitation sources coupled to 488, 532,633 and 785 nm (power 55, 50, 30 and 75 mW), all of them equipped with TruePower, as well as two high throughput spectrometers. The visible range spectrometer is equipped with 2 diffraction gratings for high and low spectral resolution (1800 and 300 lines/mm) and a CCD(EM) camera. The NIR spectrometer also has 2 diffraction gratings with 1200 and 300 lines/mm and a BI-DD CCD camera. The motorized scanning platform allows movement over an area of 50 x 50 mm2 and is coupled to a piezoelectric scanner (100 x 100 x 20 µm) that allows to obtain confocal images with a resolution close to the diffraction limit. The microscope can be equipped with temperature cell and incubator. The system is controlled with the Control SEVEN and Project SEVEN software.

Flow Cytometry Equipment

Flow Cytometer, MACSQuant Analyzer10 is a digital benchtop flow cytometer equipped with 3 lasers (405, 488 and 640nm) 10 optical channels (8 fluorescence and FCS, SSC).
Allows the acquisition in single tubes, and multiwell plates in an automatic way.
The MACSQuantify software controls sample acquisition and processing.

Optical Spectroscopy Equipment

Spectrofluorometer, Edinburgh FS5. It allows to acquire emission, excitation, and synchronous spectra from ultraviolet to near-infrared UV-NIR between 200 and 1650 nm. It also allows to record the kinetic profile at multiple wavelengths, perform polarization and fluorescence anisotropy measurements in liquid samples, thin films, and solids. It has an integrating sphere that allows measurements to be made to determine the fluorescence quantum yield and acquire diffuse reflectance spectra. The system allows the characterization of fluorescence and phosphorescence lifetimes, for this purpose it uses high intensity pulsed LEDs (340 and 405nm) and pulsed xenon lamps respectively. The system is controlled by the Fluoracle software.

Circular dichroism spectropolarimeter, CD – Jasco J-1500.
Allows measurements of absorbance, circular and linear dichroism from ultraviolet to near-infrared UV-NIR (180 and 1600 nm); It also allows to measure the fluorescence and rotational scattering fluorescence. It I equipped with a Xe light source (power 150 W), Peltier temperature control and an integrating sphere for measurements of samples with high light scattering from solids, either by reflection or by diffuse reflectance. The system is coupled to an ATS-429L automated titration system. The system is controlled with Spectra Manager II software.

Fourier Transform infrared spectrometer, FTIR – Bruker Invenio-X allows IR transmittance measurements of solid, liquid or gel samples, in the range between 30 and 15.500 cm-.1 An attenuated total reflectance (ATR) accessory equipped with diamond crystal is available and three detectors are used to cover the full range: InGaAs (NIR), DLaTGS (MIR) and DLaTGS/PE (FIR). The system is controlled by the OPUS 8.5 software. A hydraulic press is available for the compression preparation of tablets.

UV-vis-NIR spectrometers, Varian Cary 5000 allows to conduct transmittance/extinction analysis of light in solid, liquid, suspended or thin films samples, in the range between 175 and 3300 nm. The measurement system is double-beam and has a high spectral resolution (up to 0.01 nm in UV-Vis and 0.04 nm in NIR) thanks to its PMT (UV-vis) and PbS (NIR) detectors. Spectral parameters can be adjusted individually for both ranges. It is equipped with a multi-cell support that allows temperature-controlled measurements within the range between 0 and 90C, through the CaryWinUV software. It can be equipped with an integrating sphere (DRA) for diffuse reflectance measurements and an universal measurement accessory (UMA) for absolute and angle-dependent transmittance/reflectance measurements. Jasco V730 allows to measure transmittance in the range between 190 and 1100 nm for solutions and suspensions with spectral resolution of 1 nm, includes temperature control between 0-90C and magnetic stirring option. System is driven by SpectraManager 2.0.

Raman spectrometer, B&WTek i-Raman. 785 allows the analysis of macroscopic samples and materials in the form of liquids, gels, powders, and solids. The spectrometer has a laser at 785 nm (power 250 mW) as excitation source and operates in the spectral range from 175 to 3200 cm-1 with spectral resolution of 3,5 cm-1. The emitted light and the reception of the signal are obtained through an optical fibre connected to the equipment, which has a CCD detector. Signal acquisition is controlled by the BWSpec4 software.

Dynamic Light Scattering, Malvern Zetasizer Ultra Red. The combined Dynamic Light Scattering (DLS) and Electrophoretic Light Scattering (ELS) system incorporating Non-Invasive Back Scatter (NIBS) and Multi-Angle DLS (MADLS) technology enables measurements of particle or molecular size (0.3nm-10um), particle charge (for particle sizes from 4nm-100um), and particle concentration (range 10^8-10^12 particles/mL). Measurements at three angles are available (174-back, 90-side, and 13-forward). The instrument is equipped with a 10 mW 633 nm laser, fluorescence filter, vertical and horizontal polarizer, Avalanche photodiode detector and is temperature-controllable between 0-120ºC. ZS Explorer Software is used for data acquisition, analysis, evaluation, processing, and presentation.

Differential Scanning Calorimeter, Mettler DSC3+. Allows to measure crystallisation, glass transition, heat capacity of solids, gels and some liquid samples. The system is equipped with FRS5+ sensor and can be operated between -50-600ºC. STARe Software is used for data acquisition, processing and analysis.

MicroScale Thermophoresis (MST) Binding Affinity Monolith NanoTemper: Measure the strength of molecular interactions in solution, without immobilization. Based on MST measures variations in fluorescence intensity caused by ligand binding while a very precise and brief laser-induced temperature change is applied. The change in fluorescence is then plotted against the concentration of the binding partner to obtain the dissociation constant or Kd.
Can measure binding affinity data for multiple type of molecule: proteins, small molecules (ions, peptides, carbohydrates, nanoparticles), nucleic acids, vesicles, platelets and small cells, virus particles.

• Quantify low and high binding affinities, from 1nM to 500mM.
• Typical sample volume 4 μl of sample
• Excitation range Blue 450-490nm, excitation range Red 590-640nm
• Emission range Blue 505-580nm, emission range Red 660-740nm
• Molecular Weight range (Da) 101 -107