Nanbiosis

U28-S08. Cryo-TEM

Cryo-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 cryo-EM to cell biology.
• Tissue characterization: cryo-electron microscopy has a role in the characterization of interactions between cells and with other components inside tissues.
• Biomaterials characterization: cryo-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 cryo-EM.
• Macromolecular complexes characterization: structural biology can benefit from cryo-electron microscopy to determine native 3D structures of macromolecular complexes.

Target customer

Any company or research group interested in:

• Electron Microscopy of frozen-hydrated samples.
• Plunge-freezing of samples in water solution (vitrification) for cryo-transmission electron microscopy.
• Cryo-fixation of samples at high pressure (High-Pressure Freezing).
• Glow-discharge of film-coated transmission electron microscopy grids.
• Data and image processing.
• Technical advice for users regarding the selection of protocols and procedures for experiments involving electron microscopy.

References

1. Galisteo-González F, Molina-Bolívar JA, Navarro SA, Boulaiz H, Aguilera-Garrido A, Ramírez A, Marchal JA. Albumin-covered lipid nanocapsules exhibit enhanced uptake performance by breast-tumor cells. Colloids Surf B Biointerfaces. 2018 May 1;165:103-110. doi: 10.1016/j.colsurfb.2018.02.024. Epub 2018 Feb 13. PMID: 29471218.
2. Molina-Bolívar J.A., Galisteo-González F. Olive-oil nanocapsules stabilized by HSA: Influence of processing variables on particle properties. J. Nanoparticle Res. 2015;17 doi: 10.1007/s11051-015-3192-1

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

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.

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.

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.

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

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.