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OUR SERVICES

MEDFUTURE Research Center

is providing specific research services covering ‘omics’ technologies (proteomics, metabolomics),
molecular and functional imaging, in vitro and in vivo studies.

'OMICS'

‘Omics’ technologies

Shotgun proteomics technique is used for the determination of the protein profile of tissue samples.

The service includes the extraction of proteins from tissue (fresh frozen or FFPE), sample processing and analysis by nanoLC (nano-liquid chromatography) coupled with high resolution mass spectrometry.

The resulting profiles can be compared (control vs. disease), following the significantly modified proteins between the two groups.

The service includes the identification of proteins using standard databases.

Protein profiling of cell cultures lysates is determined by shotgun proteomics techniques and measured by nano-liquid chromatography coupled with high resolution mass spectrometry, after performing specific sample preparation to extract the proteins from the cell pellet.

The resulting profiles can highlight adaptations of cellular metabolism because of the treatment applied on cell cultures (treatment vs control).

The service includes the identification of proteins using standard databases.

Protein profiling of serum or plasma is determined by shotgun proteomics techniques and measured by nano-liquid chromatography coupled with high resolution mass spectrometry.

Proteome profiling can be performed on serum or plasma samples with or without depletion of the six most abundant proteins (serum albumin, immunoglobulin gamma, immunoglobulin alpha, serotransferrin, haptoglobin, alpha-1-antitrypsin), following specific sample preparation steps in order to achieve the best proteome coverage.

Specialized software are used for protein identification and relative quantification.

The analyses are done using standard or personalized microplates kit.

Microplates are read by a multiplate reader and data is analysed by a specialized software.

The analyses are performed using LC-MS untargeted methods.

After protein removal, the sample is analysed using chromatographic methods appropriate for different types of metabolites (polar/non-polar).

The resulting profiles are then compared (control vs. diseased), determining the metabolites which are significantly different among the groups.

IMAGING

Molecular and functional imaging

Acquisition of Absorbance/ Transmittance, Emission, Excitation, Excitation and Emission Matrix (EEM) and kinetics spectra.

UV-Vis-NIR Fluorescence detection domain from 250 to 1100 nm.

3D graphics (EEM) using EzSpec software can be obtained.

Automatic correction for primary and secondary inner filter effects (IFE).

The equipment can be used as a fluorimeter, UV-Vis-NIR spectrometer to measure the absorbance, or as an instrument that measures true molecular fingerprints, which require the acquisition of fluorescence, correcting for IFE in real time.

Colloids and dispersions stability analysis by measuring the zeta potential of the nanoparticles.

The system incorporates rapid pH and polyelectrolyte titrations for investigation of stable pH regions, iso-electric point, charge density and stability.

Nanoparticle’s size and concentration measurements.

The equipment can measure the hydrodynamic diameter for nanoparticles and their concentration in a solution.

The scanning is automatic, fast, and optimized by the software.

It can analyse exosomes, extracellular vesicles, liposomes, micelles, proteins, viral particles or viruses, therapeutic molecules, and fluorescently labelled nanoparticles without affecting their optical properties.

Proper for polar liquid samples (water, alcohol) or organic solvents.

Lasers: 375nm, 405nm, 488nm, 520nm, 640nm, 660nm.

TEM analysis of different types of nanoparticles and nanostructures.

This analysis includes sample preparation and image acquisition.

TEM analysis of biological samples for ultrastructure analysis.

The protocol includes sample preparation (prefixation, post-fixation, dehydration, resin infiltration and embedding, resin block modeling, semi-fine sectioning, staining, optical analysis, resin block remodeling, ultra-thin sectioning, counter-staining) and image acquisition.

Spectral analysis of different types of samples (solid/liquid/gel).

Excitation wavelength interval from UV to NIR using 5 lasers (325, 532, 633, 785 and 830 nm).

The Raman protocol involves the deposition of the sample on aluminium foil or CaF2 glass sample holder.

The SERS protocol involves the use of a new type of solid plasmonic substrate developed in our laboratory using TFF processed silver nanoparticles.

Topographic imaging at nanometric resolution for solid surfaces.

Suitable for nanoparticles and biological samples.

Imaging of cultured cells, fluorescent molecules, fluorescently labelled cellular structures.

The protocol can include fluorescence labelling using fluorophores or primary antibodies and secondary fluorescently labelled antibodies for immunostaining.

The equipment allows the analysis of a sample stained with three different dyes.

Lasers: 405, 488, 515, 543, 633 nm.

Label-free Darkfield imaging of plasmonic nanoparticles and/or biological samples.

It allows the analysis of cellular and subcellular structures, morphological differences between treated and untreated cells, cellular internalization of nanoparticles.

IN VITRO

In vitro studies

Bioactive compounds testing includes cell culturing steps (cell subculturing and biobanking) using cells from the research centre biobank or purchased from a third-party.

The cytotoxicity of these compounds is tested using colorimetric assays (MTT, XTT, MTS) and the samples are analysed by microplate reader.

Protein quantification can be performed using the NanoDrop device or by classical protein dosing methods (Bradford, BCA).

The WesternBlot step itself includes the migration and transfer of migrated proteins through the electrophoresis system, and the labelling of proteins with specific antibodies for each protein.

The samples are analysed with the chemiluminescent detection device.

Determination of gene expression by RT-PCR includes the steps of extracting the genetic material using TRIZOL or the DNA / RNA extraction and purification kit.

Also, it includes the steps of cDNA synthesis and the chain polymerization reaction itself.

The sample read is performed on StepOnePlus PCR Machine.

IN VIVO

In vivo studies

The service includes the accommodation of small animals (mice and rats) with immunocompromised phenotype or with artificially modified genotype (transgenic animals) in conditions of strict sterility, with ad libitum autoclaved water and food intake and autoclaved bedding change on a biweekly basis.

The service also includes daily monitoring of animal health and behaviour by authorized personnel and the administration of supportive treatments for mild pathological conditions.

If the animal’s health is incompatible with life, the researcher will be notified, and the animal will be sacrificed after 24 hours (the service also includes the collection of organs in the case of a requested necropsy).

Bioluminescence monitoring of the distribution of cells or molecules/particles in immunocompromised mice involves the injection of genetically transformed systems that express the enzyme luciferase, systems that can be tracked through IVIS equipment after systemic administration of the specific substrate – luciferin.

The service includes the preparation of the experimental animal according to the specific protocol, administration of the bioluminescent substrate, administration of gas anaesthesia, imagistic evaluation, data acquisition and processing, including support in data interpretation, and follow-up of the experimental animal until recovery.

Fluorescence monitoring of the distribution of cells or molecules/particles in immunocompromised mice involves the administration of fluorescence transformed/conjugated systems, systems that can be tracked through IVIS equipment.

The service includes the preparation of the experimental animal according to the specific protocol, administration of fluorescent systems, administration of gas anaesthesia, imagistic evaluation, data acquisition and processing, including support in data interpretation, and follow-up of the experimental animal until recovery.

The service includes pre-operative preparation of animals that includes the establishment of an aseptic field, gas/injectable anesthesia services, the actual intervention, post-operative recovery and monitoring.

The staff of the Animal Facility Department has supra-specialization trainings that allow interventions such as: vasectomy, adrenalectomy, nephrectomy, ovariectomy, ovarian transplantation, jugular vein catheterization, development of orthotopic models for malignant pathologies, procedures adapted to the researcher’s requirements.

The service includes the preparation of biological samples for haematological determinations, the performance of determinations using an equipment dedicated to veterinary use and the transmission of results.

The service includes the handling of rodents, the performance of procedures, the post-procedural monitoring of animals and the proper storage of the sample until delivery/processing.