JRC

Name of the infrastructure: JRC-TA

Location (town, country): Ispra (Varese), Italy

Web site address:

Legal name of organisation operating the infrastructure: Joint Research Centre of the European Commission

Location of organisation (town, country): Brussels, Belgium

Description of the Infrastructure

MATERIALS AND EXPERTISEThe Joint Research Centre is a large multinational centre that carries out research in a wide variety of fields,including Health, Environment, Energy, and Safety/Security of European citizens. This offers users of thelaboratories opportunities for establishing new contacts and for novel multidisciplinary research activities. TheInstitute for Health and Consumer Protection (IHCP) itself is one of the 3 scientific institutes at the JRC Isprasite. The other two are the Institute for Environment and Sustainability (IES) and the Institute for Protectionand Security of the Citizen (IPSC). Altogether about 1700 permanent and temporary staff from all 27 EUmember states, as well as several associated countries, work at the JRC Ispra site. This makes Ispra one ofthe unique large multicultural research centres in Europe, providing a highly stimulating environment for visitingresearchers. The IHCP has several laboratories involved in research on Nanotechnology and Nanomaterials, theNanobiotechnology Laboratory, the Cyclotron Laboratory, and the High Throughput System (HTS) Laboratory.These are hosted in two Units – the Nanobiosciences (NBS) Unit, and the Systems Toxicology (ST) Unit. In theNBS and ST Units of IHCP, about 100 scientists, technicians and research students are working on the areasof research outlined above. In the other 3 Units of IHCP, another 150 are working on methods of detection ofGMO in food products, on indoor air pollution and on the safety of chemicals. The JRC has been involved inNanotoxicity work for the past 5 years and has developed a strong competence in NanoBiotechnology, and inparticular in Nanoparticles synthesis, labelling and functionalisation, materials characterisation, cell interactionand in vitro toxicity testing and protein surface interactions.Apart from an institutional workprogramme focused mainly on research into safety of nanoparticles, the JRC isinvolved in several competitive projects and collaborations on this subject. It has been strongly engaged in thedevelopment of international programmes related to Nanotoxicology, in particular by co-chairing several workinggroups in the OECD Working Party on Manufactured Nanomaterials, as well as CEN and ISO specialisedgroups. It has participated in several FP6 and FP7 competitive projects such as DIPNA, CellNanoTox, IntelTex,Nanother, Nanotest, Nanoretox and NHECD.

Specific research areas include:

• Development of protocols for in vitro toxicity testing of engineered nanoparticles
• Development of nano-engineered surfaces for control of protein-surface and cell surface interactions
• Development of nanoparticle complexes for cell sensing and imaging (Quantum Dots and related core shellstructures)
• Development of methods for detection of nanoparticulate material in consumer products
• Research into protein-surface interactions and protein misfolding (NMR, Mass Spectrometry)
• Research into the synthesis of radioactive nanoparticles for eventual application in medical therapy
• Development of cyclotron techniques for radiolabelling of engineered nanoparticles.
• In vitro cellular uptake studies using radiolabelled nanoparticles.
• Research into cell based assays for toxicity testing using automation technologies and a High ThroughputScreening (HTS)format
• Development of High Content Analysis (HCA) in vitro assays using dedicated platforms for automated imagingand electrophysiological measurements
• Development of protocols for in vitro toxicity testing using label-free detection techniquesIn addition to these areas that are specifically aimed at nanotechnology and nanosafety, several other researchprojects are carried out, including development of plasma methods for surface decontamination and sterilisation,development of NPs for cell imaging and nanomedicine, and investigations into the cyclotron production of novelisotopes for application in Nuclear Medicine.For TA users, the range of competences available within the institute will represent an excellent opportunity.TNA will be offered mainly for advanced research work on the radiolabelling of nanoparticles and forcharacterisation of radiolabelled and non-labelled NPs.

SUPPORTING FACILITIES

Particle synthesis and labelling:

Two JRC laboratories are involved in nanoparticle synthesis. The Nanobiotechnology Lab. has a chemistrylaboratory specialised in the synthesis of NPs, together with 150 m2 of class 1000 clean room with stateof the art equipment. The Cyclotron Lab. has chemical hoods and glove boxes suitable for radiochemicalsynthesis of radiolabelled NPs and/or manipulation of directly activated NPs. In QNano, all TNA access toJRC facilities will be for projects that focus on the use of the Cyclotron Laboratory, specifically for synthesisand characterisation of radiolabelled NPs. Access to the Nanobiotechnology Laboratory will be offered forpreliminary synthesis studies prior to user “hands-on” synthesis studies with radioactive materials. It is notplanned to provide non-radioactive NPs to external users as TA. The combination of radiochemical synthesisfacilities, cyclotron isotopes, and NP characterisation instruments within the cyclotron controlled area, togetherwith expertise and an extensive range of instrumentation for preliminary synthesis experiments in the associatedNanobiotechnology Lab. is quite unique in Europe. These facilities are supported by 3 permanent scientists and 3 technical staff (permanent).

The main facility at the JRC for nanoparticle labelling is the Cyclotron Laboratory which hosts a ScanditronixMC40 cyclotron accelerator, the commissioning of which was completed in 1982. It is one of a very limitednumber of such research accelerators in Europe, and the only one deeply involved in nanoparticle activationresearch. The MC40 is a variable energy machine that can accelerate a range of light ions – protons, alphas,deuterons and 3He up to energies of 39MeV (for protons), up to a reasonably high current of 60￿A. It istherefore ideal for studying and applying a wide range of nuclear reactions for different applications. On onebeamline a novel neutron activator has been installed, also allowing materials activation by neutron irradiation.Apart from the cyclotron itself, the Cyclotron Laboratory offers a range of instrumentation for characterisationof radiolabelled NPs, as described below. Both the neutron activator and a dedicated ion target system areavailable for nanoparticle activation, and the instrumentation and on-site expertise in radiolabelling of NPs bydirect activation is unique. These facilities are supported by 4 permanent scientists and 4 technical staff.

Particle characterisation:

For particle characterisation, the Nanobiotechnology Lab. has a wide range of equipment available, togetherwith extensive on-site expertise in the characterisation of nanoparticles and of nano-objects: a Focused IonBeam/Field Emission Scanning Electron Microscope (FEI Nova), SEM/EDX (Leo), X-Ray Photo-electronSpectroscopy (Kratos Axis Ultra), a Time of Flight SIMS (Ion-Tof IV with a Bi gun), Dynamic Light Scattering andZeta potential (Malvern ZetaS-nano), Particle Tracking Analysis (NanoSight), Fluorescence spectrophotometer(Varian Eclipse), FTIR with liquid cell (Brucker Tensor 27), Surface Plasmon Resonance imaging (Genopics),AFM and SNOM (Agilent), Spectroscopic Ellipsometry Imaging (Lot Oriel, Nanofilm), Raman SpectroscopyImaging (Reinishaw), Optical microscopy (Zeiss, Olympus).Limited access to such instrumentation will onlybe provided if necessary, and normally only for studies related to nanoparticle radiolabelling. Access has tobe discussed and agreed, and can only be offered if scientific and technical support is available during thescheduled TNA access. In the cyclotron controlled area the main equipment offered for TNA is located. Thisincludes calibrated gamma and alpha spectroscopy systems are available, together with a grazing incidenceXRD system (suitable for NP characterisation) and a second Malvern ZetaS-nano (Dynamic Light Scatteringand Zeta potential) for characterisation of radioactive nanoparticles.Access to these instruments, as well as thecyclotron itself for NP radiolabelling will be fully available for TNA and approved TNA projects will be scheduled as far as possible to match the user’s requests. These instrumentations are supported by 6 permanent scientistsand 6 technicians.

Research supported by the infrastructure

The JRC laboratories are involved in several funded collaborative research projects that include nanoparticlesynthesis, characterisation, and in vitro testing. The various facilities and instruments are linked to external usersmainly through these projects, though in several cases external users directly access the laboratories for theirwork with scientific and technical support provided by JRC sTAf. The specific projects can be grouped under thefollowing components:

• Particle synthesis and functionalisation: Synthesis of a large range of nanoparticles (gold, cobalt ferrite,SiO2, Ag, ZnO, CeO2, ZnS, CdSe) by physical and chemical methods with controlled sizes and physicochemical properties. Production of core shell structures for cellular trafficking and imaging. Direct synthesis ofradiolabelled nanoparticles from radioactive precursor materials.
• Particle Characterisation: done on as prepared materials (size distribution, surface charge, composition,crystallographic structure and shapes etc…) and after interaction with biological relevant medium. Study ofNP-protein interaction kinetics with SPR, DLS, Field Flow Fractionation- MS. Protein conformation changes arestudied by Raman Confocal Microscopy, FTIR and NMR. Advanced Omics technologies including NMR and MSfor study of NP- protein interactions.