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U28-S07. Transmission Electron Microscopy (TEM)

Transmission Electron Microscopy (TEM)

The Electron Microscopy Service offers access to transmission electron microscopy (TEM), cryo-TEM and electron tomography, high resolution scanning electron microscopy (SEM), and environmental SEM. A special emphasis has been made on high-end sample preparation techniques through cryo-immobilization. All the equipment has been configured to provide its best for biological and nanomedicine applications.

Customer benefits

We have SOPs and ISO9001 certification. We also have specialist technicians for the use of the equipment.

This service is essential to:

  • Cellular characterization: cell structures and ultrastructures visualization and its relationship to function are the most important contributions of EM to cell biology.
  • Tissue characterization: electron microscopy has a role in the characterization of interactions between cells and with other components inside tissues.
  • Biomaterials characterization: electron microscopy plays a double role in biomaterial characterization On one hand, it provides structural and compositional information on the materials engineered to be used inside biological systems, favouring its development On the other hand, it allows us to visualize their interactions Functionalized polymeric and metallic nanoparticles for drug delivery, dental implants, bone plates and cements, and artificial tissues are among the biomaterials that can benefit from the information provided by EM.
  • Macromolecular complexes characterization: structural biology can benefit from electron microscopy to determine 3D structures of macromolecular complexes.
  • Negative staining is also a very valuable tool for 2D-3D characterization.

Target customer

Any company or research group interested in:

  • Transmission electron microscopy of resin-embedded and non-embedded samples.
  • Cryo-fixation of samples at high pressure (High-Pressure Freezing).
  • Resin embedding of samples at room temperature (conventional) or low temperature (freeze-substitution) for ultramicrotomy.
  • Ultramicrotomy of resin-embedded sections (semithin and ultrathin sections).
  • Data and image processing.
  • Technical advice for users regarding the selection of protocols and procedures for experiments involving electron microscopy.

References

  1. Gómez-González E, González-Mancebo D, Núñez NO, Caro C, García-Martín ML, Becerro AI, Ocaña M. Lanthanide vanadate-based trimodal probes for near-infrared luminescent bioimaging, high-field magnetic resonance imaging, and X-ray computed tomography. J Colloid Interface Sci. 2023 Sep 15;646:721-731. doi: 10.1016/j.jcis.2023.05.078. Epub 2023 May 18. PMID: 37229990.
  2. Caro C, Guzzi C, Moral-Sánchez I, Urbano-Gámez JD, Beltrán AM, García-Martín ML. Smart Design of ZnFe and ZnFe@Fe Nanoparticles for MRI-Tracked Magnetic Hyperthermia Therapy: Challenging Classical Theories of Nanoparticles Growth and Nanomagnetism. Adv Healthc Mater. 2024 Feb 2:e2304044. doi: 10.1002/adhm.202304044. Epub ahead of print. PMID: 38303644.

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U28-S06. Time Domain NMR (TD-NMR)

Time Domain NMR (TD-NMR)

The NMR service is equipped with a TD-NMR analyzer for relaxivity measurements at low magnetic field (1.44 T).

Customer benefits

We have SOPs and ISO9001 certification. We also have specialist technicians for the use of the equipment.

This service is essential for relaxivity measurement of magnetic compounds, such as magnetic nanoparticles or compounds containing lanthanides (Gd, Dy, etc.)

Target customer

Any company or research group interested in relaxivity measurements (r1 and r2) at low magnetic field (1.44 T).

References

  1. Caro C., Paez-Muñoz J.M., Beltrán A.M., Leal M.P., García-Martín M.L. PEGylated Terbium-Based Nanorods as Multimodal Bioimaging Contrast Agents. ACS Appl. Nano Mater. 2021;4:4199–4207. doi: 10.1021/acsanm.1c00569.
  2. Ashish Avasthi, Carlos Caro, María Luisa Garcia-Martin, Manuel Pernia Leal. Deciphering the parameters to produce highly reproducible and scalable iron oxide nanoparticles. Reaction Chemistry & Engineering 2023, 8 (7) , 1638-1653. doi: 10.1039/D2RE00516F
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U28-S05. High Resolution Magic Angle Spinning (HR-MAS) NMR

High Resolution Magic Angle Spinning (HR-MAS) NMR

The NMR service is equipped with a High resolution NMR Spectrometers: a 600 MHz, spectrometer dedicated NMR studies that are more demanding in terms of sensitivity. This system is also equipped with a high resolution magic angle spinning (HR-MAS) NMR to analyze heterogeneous samples (tissue biopsies or partially insoluble compounds).
Applications of NMR spectroscopy to chemistry range from structure elucidation of small molecules to studies of ligand-protein interactions.

Customer benefits

We have SOPs and ISO9001 certification. We also have specialist technicians for the use of the equipment.

This service is essential for metabolomic analysis of intact tissue (biopsies)

Target customer

Any company or research group interested in:

  • Acquisition of 1 H and 31P HR-MAS NMR spectra.
  • Preparation of tissue samples for HR-MAS NMR spectroscopy.
  • Basic processing of NMR spectra.

References

Kawashima Y, Nagai H, Konno R, Ishikawa M, Nakajima D, Sato H, Nakamura R, Furuyashiki T, Ohara O. Single-Shot 10K Proteome Approach: Over 10,000 Protein Identifications by Data-Independent Acquisition-Based Single-Shot Proteomics with Ion Mobility Spectrometry. J Proteome Res. 2022 Jun 3;21(6):1418-1427. doi: 10.1021/acs.jproteome.2c00023. Epub 2022 May 6. PMID: 35522919; PMCID: PMC9171847.

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U28-S04. High Resolution Liquid NMR

High Resolution Liquid NMR

The NMR service is equipped with a High resolution NMR Spectrometers :a 400MHz (NMR spectrometer for routine experiments and a higher filed) y 600MHz (spectrometer dedicated NMR studies that are more demanding in terms of sensitivity)
Applications of NMR spectroscopy to chemistry range from structure elucidation of small molecules to studies of ligand-protein interactions. In biomedicine, the use of NMR spectroscopy has expanded from the classical metabolic studies using enriched stable isotope tracers to the more recent NMR-based metabolomics of biofluids (urine, plasma, CSF).

Customer benefits

We have SOPs and ISO9001 certification. We also have specialist technicians for the use of the equipment.

This service is essential to:

  • Structure verification and elucidation
  • Molecular mobility: determination of conformation in solution
  • Determination of enantiomeric excess
  • Kinetic and temperature studies of reaction mixtures
  • Metabolomic analysis of biofluids (urine, blood serum, etc.)

Target customer

Any company or research group interested in:

  • Acquisition of 1D NMR spectra at 400 MHz and 600MHz (1 H, 13C, 31P, etc.)
  • Acquisition of basic 2D experiments: COSY, TOCSY, HSQC, HMBC, J-RES
  • Nuclear Overhauser Effect experiments (NOE and ROE), Diffusion experiments (DOSY), selective excitation experiments.
  • Preparation of biofluid samples (urine, blood serum/plasma) for high resolution NMR spectroscopy following standardized protocols.
  • Basic processing of NMR spectra.

References

  1. Pomary PK, Eichau S, Amigó N, Barrios L, Matesanz F, García-Valdecasas M, Hrom I, García Sánchez MI, Garcia-Martin ML. Multifaceted Analysis of Cerebrospinal Fluid and Serum from Progressive Multiple Sclerosis Patients: Potential Role of Vitamin C and Metal Ion Imbalance in the Divergence of Primary Progressive Multiple Sclerosis and Secondary Progressive Multiple Sclerosis. J Proteome Res. 2023 Mar 3;22(3):743-757. doi: 10.1021/acs.jproteome.2c00460. Epub 2023 Jan 31. PMID: 36720471; PMCID: PMC9990127.
  2. Molina N, Martin-Serrano A, Fernandez TD, Tesfaye A, Najera F, Torres MJ, Mayorga C, Vida Y, Montañez MI, Perez-Inestrosa E. Dendrimeric Antigens for Drug Allergy Diagnosis: A New Approach for Basophil Activation Tests. Molecules. 2018 Apr 24;23(5):997. doi: 10.3390/molecules23050997. PMID: 29695102; PMCID: PMC6100007.
  3. Fernández-Palacios S, Matamoros E, Morato Rojas I, López Navarrete JT, Ruiz Delgado MC, Vida Y, Perez-Inestrosa E. New Insights into Acylhydrazones E/Z Isomerization: An Experimental and Theoretical Approach. Int J Mol Sci. 2023 Sep 29;24(19):14739. doi: 10.3390/ijms241914739. PMID: 37834186; PMCID: PMC10648745.
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U28-S03. Computed Tomography (CT)

Computed Tomography (CT)

Computed Tomography (CT) is based on the reconstruction of 2D X-ray images captured at different angles to generate a detailed 3D view of the subject. CT imaging is typically used to provide vital contextual information for other modalities as well as for specific studies into bone density, contrast agents, implantation and regenerative technologies. Our Albira system uses dedicated beds for rats and mice with incorporated gas anesthesia. The Albira software suite automates the reconstruction process, while the PMOD software suite enables advanced analyses and the export of data into popular formats such as DICOM.

Customer benefits

We have SOPs and ISO9001 certification. We also have specialist technicians for the use of the equipment.

This service is essential to:

  • Dentistry and bone regeneration studies
  • Contextual/anatomical imaging in conjunction with other modalities
  • Bone density analysis
  • Implantation studies

Target customer

Any company or research group interested in:

  • Phantom-based measurement of reagent densities (e.g. as part of the characterization of X-ray/CT-based contrast agents)
  • Bone-density measurements based on calibration using a calcium hydroxyapatite phantom.
  • Image processing, analysis and quantification services, including specialized packages such as PMOD, FIJI and BoneJ.
  • Advanced custom 3D visualizations using the Imaris software package.
  • Close integration with other BIONAND units for associated animal handling and histology services.

References

  1. Caro C, Gámez F, Quaresma P, Páez-Muñoz JM, Domínguez A, Pearson JR, Pernía Leal M, Beltrán AM, Fernandez-Afonso Y, De la Fuente JM, Franco R, Pereira E, García-Martín ML. Fe3O4-Au Core-Shell Nanoparticles as a Multimodal Platform for In Vivo Imaging and Focused Photothermal Therapy. Pharmaceutics. 2021 Mar 20;13(3):416. doi: 10.3390/pharmaceutics13030416. PMID: 33804636; PMCID: PMC8003746.
  2. Christou E, Pearson JR, Beltrán AM, Fernández-Afonso Y, Gutiérrez L, de la Fuente JM, Gámez F, García-Martín ML, Caro C. Iron-Gold Nanoflowers: A Promising Tool for Multimodal Imaging and Hyperthermia Therapy. Pharmaceutics. 2022 Mar 14;14(3):636. doi: 10.3390/pharmaceutics14030636. PMID: 35336012; PMCID: PMC8955043.
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U28-S02. Fluorescent and Bioluminescent Optical Imaging

Fluorescent and Bioluminescent Optical Imaging

The In-Vivo Xtreme is a multi-modal optical and X-ray small animal imaging system, designed to combine sensitivity, speed and flexibility for the broadest range of applications for molecular imaging. Equipped with a high performance f/1.3 58mm lens and 4MP back-illuminated CCD camera, the In Vivo Xtreme is suitable for demanding bio-luminescence and fluorescence intravital imaging challenges

Customer benefits

We have SOPs and ISO9001 certification. We also have specialist technicians for the use of the equipment.

This service is essential to:

  • Bioimaging of labeled cells/tissues using fluorescence or bioluminescencee
  • Monitoring of Drug Delivery and Targeting
  • Pharmacodynamic studies
  • Imaging and validating new Probes and Biomarkers

Target customer

Any company or research group interested in:

  • Phantom-based experimental assessment of labeling strategies
  • Real-time bioimaging of labeled cells or pharmacological agents during and immediately after intravenous injection
  • Medium-term (1-14 days) biodistribution studies of fluorescently-labeled agents over days or weeks with ex-vivo validation
  • Design custom spectral unmixing profiles to reduce autofluorescence or crosstalk between overlapping markers
  • Establishment of disease models using commercial molecular imaging reagents
  • Close integration with other BIONAND units for associated animal handling and histology services
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U28-S01. Magnetic Resonance Imaging (MRI)

Magnetic Resonance Imaging (MRI)

Magnetic Resonance Imaging (MRI) is a very versatile technique that offers not only excellent morphological information, but also functional and physiological. Furthermore, important advances during last years in the development of highly sensitive contrast agents using nanotechnology approaches have brought MRI into the molecular imaging area. Thus, small animal MRI scanners have become a key tool in the development of magnetic nanoparticles for medical diagnosis and therapy.

Customer benefits

We have SOPs and ISO9001 certification. We also have specialist technicians for the use of the equipment.

This service is essential to:

  • Anatomical Studies: tumors, inflammation, Degenerative diseases, etc.
  • Functional Studies: brain perfusion, vascular permeability, in vivo metabolic profiling, etc.
  • Pharmocokinetics and Biodistribution of new Contrast Agents (or any drug magnetically labeled)
  • Monitoring of Drug Delivery and Targeting
  • Follow up of Therapy Response

Target customer

Any company or research group interested in:

  • Morphological imaging: T1-weighted, T2-weighted, PD, diffusion-weighted, etc.
  • T1 and T2 Dynamic Imaging.
  • Diffusion Imaging: ADC, DTI.
  • Phantom-based relaxivity measurements.
  • Cardiac Imaging.
  • Single Voxel Spectroscopy.
  • Image processing, analysis, and quantification.
  • Close integration with other BIONAND units for associated animal handling and histology services.

References

  1. Caro C, Guzzi C, Moral-Sánchez I, Urbano-Gámez JD, Beltrán AM, García-Martín ML. Smart Design of ZnFe and ZnFe@Fe Nanoparticles for MRI-Tracked Magnetic Hyperthermia Therapy: Challenging Classical Theories of Nanoparticles Growth and Nanomagnetism. Adv Healthc Mater. 2024 Feb 2:e2304044. doi: 10.1002/adhm.202304044. Epub ahead of print. PMID: 38303644.
  2. Caro C, Avasthi A, Paez-Muñoz JM, Pernia Leal M, García-Martín ML. Passive targeting of high-grade gliomas via the EPR effect: a closed path for metallic nanoparticles? Biomater Sci. 2021 Nov 23;9(23):7984-7995. doi: 10.1039/d1bm01398j. PMID: 34710207.
  3. Borrego LG, Recio R, Moreno N, Chelouan A, Álvarez E, Sánchez-Coronilla A, Caro C, Pearson JR, García-Martín ML, Khiar N, Fernández I. Enantioselective synthesis of 4-amino-3,4-dihydrocoumarins and their non-cyclic hydroxyester precursors: Biological evaluation for the treatment of glioblastoma multiforme. Eur J Med Chem. 2022 Dec 5;243:114730. doi: 10.1016/j.ejmech.2022.114730. Epub 2022 Sep 1. PMID: 36088758.
  4. Gómez-González E, Núñez NO, Caro C, Garcia-Martín ML, Ocaña M. Carboxylate functionalized NaDy(MoO4)2 nanoparticles with tunable size and shape as high magnetic field MRI contrast agents. J Colloid Interface Sci. 2023 Jan;629(Pt A):310-321. doi: 10.1016/j.jcis.2022.08.130. Epub 2022 Aug 24. PMID: 36081210.
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STRYKER & NANBIOSIS start a promising line of biomedical research collaboration

On January 23-24th a two-day meeting between NANBIOSIS and Stryker Trauma GmbH, took place at the Carlos III Health Institute, in Madrid, to explore synergies and potential joint initiatives.  

Nils Reimers and Robin Buescher from Stryker, presented some of their research and upcoming projects in the field of trauma medical devices, and representatives from NANBIOSIS’ units -Marisa González from U16 Surface Characterization and Calorimetry Unit, Teresa Galán from U7, Nanotechnology Unit, Gemma Pascual from U17 Confocal Microscopy Service, from CIBER-BBN, Marisa García from U28 (Bionand) and José Francisco Guillén and Francisco Sanchez Margallo from the units of the Jesús Usón Minimum Invasive Surgery Center (CCMIJU) , together with Jesús, Izco Executive Director of NANBIOSIS and Ramon Martínez, Scientific Director of CIBER-BBN- explained their technologies.

The second day, the meeting location was moved to the CCMIJU in Cáceres, to visit the Center facilities and continue conversations.

All attendees expressed their satisfaction with the development of the meeting and agreed to continue with the explorations in forthcoming meetings.

Photo caption: From left to right: José Francisco Guillén (Quality Guarantee Manager of CCMIJU),  Jesús Izco (NANBIOSIS Executive Director), Francisco Sanchez Margallo (Scientific Director of CCMIJU), Nils Reimers (Senior Manager R&D – Stryker Trauma GmbH), Robin Buescher (Director R&D Stryker Trauma GmbH), Margarita Casado (Management Staff of NANBIOSIS)”

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Laura Lechuga, Scientific Director of Unit 4 of NANBIOSIS Coordinates a EuroNanoMed project

ABISENS Monitoing of Acquired Brain Injury and recovery biomarkers by the combined label-free nanosensing of multiple circulating molecules is one of the project awarded in the EuroNanoMed III joint call, will be coordinated by Laura Lechuga through the CIBER-BBN

Abisens will employ the Unit 4 of Nanbiosis, Biodeposition and Biodetection Unit or the multiplexed biofunctionalization of the biosensor chips and their methodology optimisation

The evaluation of patients after brain injuries, which produces severe impairments, remains a major unmet clinical need. Nowadays the diagnosis, prognosis and the efficacy of rehabilitation treatments are mainly assessed by clinical examinations, neuroimaging and electrophysiological tests during a long hospitalization stay. The aim the project is to offer as an alternative a new nanobiosensor platform able to identify and quantify multiple brain biomarkers in blood with high sensitivity and in a short time. The new biosensor platform will employ nanophotonic waveguide circuits in combination with oligonucleotide chemistry.

The project will be characterized by a strong interdisciplinary and translational nature resulting from the meeting between real clinical needs and high-level technological integration of biosensing and bimolecular aspects.

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NANBIOSIS Against Cancer

The World Health Organization, the International Cancer Research Center (IARC) and the International Union Against Cancer (UICC) celebrate February 4 of each year as World Cancer Day

Every year, 14 million new cases of cancer are diagnosed worldwide and the disease causes 8.2 million deaths.

Thanks to scientific research, great advances have been made in the fight against cancer. Through surgery, chemotherapy or radio therapy and, in the last 20 years, through immunotherapy, hormonal treatment or cell therapies, tools have been obtained to improve early diagnosis and treatments, increasing cancer survival by 20%.

The only way to understand cancer and, someday, eradicate it or eliminate the suffering and death due to this disease, is RESEARCH

NANBIOSIS as an ICTS (Singular Scientific and Technical Infrastructures) for biomedical research plays a very important role in the fight against cancer. Some examples are bellow:

Thanks to a coordinated action between units U1 of Protein Production Platform (PPP), U18 of Nanotoxicology and U29 of Nucleic Acid Synthesis, NANBIOSIS is developing nanopharmaceuticals with a high degree of efficacy for the treatment of metastases in colon cancer, by using of proteins with high specificity of binding to metastatic cells and a high degree of permanence in the blood flow, loaded with anti-cancer drugs that are selectively released inside the tumor cells that are going to form the metastases. Through the public financing of a NEOTEC project and a RETOS-COLABORACION and the company NANOLIGENT SL, the first antimetastatic drug on the market will be developed.

The Protein Production Platform-PPP collaborates with research projects whose objective is the development of new cancer therapies based on recombinant modular proteins with the ability to self-assemble. These multimeric complexes have shown, in animal models, a high stability in serum and an improved biodistribution compared to that observed with drugs for clinical use. These principles have been valued in different types of cancer, including colorectal cancer and breast cancer. The modular design of these constructions allows the incorporation or substitution of direct peptides and therefore they are presented as a transversal tool for more effective treatments against cancer. In addition, the PPP has served the Vall d’Hebron Institute of Oncology (VHIO) of Barcelona, the Josep Vilanueva group (CIBERONC) in the field of biomarker study and new targets associated with triple negative breast cancer (TNBC).

Unit 6 of NANBIOSIS Biomaterial Processing and Nanostructuring Unit is working on a project in collaboration with VHIR, financed by the Spanish Goverment and CIBER-BBN, for the development of a new nanomedicine for the treatment of high-risk neuroblastoma, one of the most frequent childhood cancers.

Unit 6 is also working on the project Artificial Lymph Nodes for Cancer ImmunoTherapy (ALYCIA) A project born of a initiative of CIBER-BBN/ CIBERONC to enhance scientific interdisciplinary collaborations between research groups working on oncology and nanomedicine. Researchers of unit 6 will develop Artificial Lymph Nodes (ALN) based on dynamic 3D scaffolds able to promote efficient ex vivo lymphatic cell expansion of relevant phenotypes. Such ALN represent a new approach to lymphocyte expansion, which not only includes artificial Antigen Presenting Cells in suspension like the state-of-the-art expansion techniques, but also mimics the function of the LN ex vivo.

One of the singular capabilities of the U25 of NANBIOSIS NMR: Biomedical Applications I is the acquisition of high quality, high resolution preclinical magnetic resonance imaging/spectroscopy/spectroscopic imaging data. This allows performing leading-edge studies in preclinical cancer models such as noninvasive therapy response follow-up in murine brain tumours, revealing new response biomarkers with translational potential for brain cancer patients.

NANBIOSIS U4 Biodeposition and Biodetection Unit  is currently developing the national project PREDICT Point-of-care Nanoplasmonic Platforms for Novel High-Value Diagnostics and Therapy Follow-Up , which works in the early detection of lung cancer. PREDICT project will use the Unit 4 of Nanbiosis for the multiplexed biofunctionalization of the biosensor chips and their methodology optimisation.

Finally, Unit 20 of NANBIOSIS In Vivo Experimental Platform at VHIR, is the most implicated of the CIBER units on projects in the field of cancer, just to name some of them: H2020-NoCanTher: magnetic nanoparticles against pancreatic cancer through the use of hyperthermia combined with conventional treatment. H2020-Target-4-Cancer: nanotherapy based on polymeric micelles directed against specific receptors of tumor stem cells in colorectal cancer. H2020-DiamStar: nanodiamonds directed against leukemia for the potentiation of chemotherapy. FET-OPEN EvoNano: in silico and tumor-tumor models for the prediction of PK / PD and tumor efficacy of antitumor nanomedicines against tumor stem cells. FIS-ISCIII: polymeric micelles for siRNA and combined therapy against breast cancer tumor stem cells. CarboXigel: hydrogels for the sustained release of chemotherapeutic drugs against the metastatic spread of ovarian cancer. MelanoMir: nanomedicine applied to skin cancer, melanoma, beside other projects promoted by CIBER-BBN.

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