LabRMN-CNC – Nuclear Magnetic Resonance Laboratory
Center of Neurosciences and Cell Biology, University of Coimbra
The laboratory is currently part of the Area E “Biophysics and Biomedical NMR” of the CNC. It is also part of the Portuguese National Network of Nuclear Magnetic Resonance Spectroscopy (PTNMR), under the supervision of the Foundation for Science and Technology (FCT), Portugal.

The main objectives of the LabRMN-CNC are the provision of Nuclear Magnetic Resonance to the scientific community, the disclosure and promotion of NMR and training of users. In addition of developing its own research projects, the LabRMN-CNC aims to support work of scientists of the national academic community as business environment, from public or private institutions, thru collaboration in research projects or provision of services.

NMR Unit Coordinator:
Prof. Carlos F.G.C. Geraldes, PhD
Contact: geraldes@bioq.uc.pt

NMR Unit Manager:
Dr. Emeric Wasielewski, PhD
Contact: emeric@cnc.uc.pt
Voice: +351 304 502 926
Fax:     +351 239 853 409

NB: Meeting with the NMR Unit Coordinator or the NMR Unit Manager are with appointment only. Appointments can be requested per E-mail at least 24 hours in advance.
NMR Time schedule:
NMR time schedules are planned on a monthly basis, beginning the first Monday of the month.

Online calendars Periods
Red: NMR events (as planning days) M => morning (9:30 AM to 1:45 PM)
Mustard: VNMRS 600 MHz schedule A => afternoon (1:45 PM to 6 PM)
Aluminium: Unity 500 MHz schedule D => daytime (9:30 AM to 6 PM)
Cyan: Open time (for both spectrometer) N => night (6 PM to 9:30 AM)
Violet: Portuguese holidays. F => full day (start at 9:30 AM)
Additional information about session is available by clicking on the name.

NB: For more information about the operating of the LabRMN-CNC, please read the "Guide for NMR access and use" below.
Research Interests:
A. Inorganic Complexes for Medical Diagnostics: MRI and Molecular Imaging:
Studies focus on inorganic compounds for medical diagnostic imaging, in particular contrast agents for magnetic resonance imaging (MRI CAs). They include design, development and physico-chemical evaluation of metal based nanoparticles and small metal chelate agents for multimodal targeted medical imaging agents, followed by in vitro and animal model evaluation using MRI and nuclear imaging techniques.
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B: Inorganic Biochemistry: Inorganic Complexes and Species for Therapy and Environment:
We are interested on the design and study of inorganic compounds as drugs for medical therapy (lithium Li+ and bipolar disorder; vanadium V(IV/V) complexes as oral insulin-mimetic agents). We focus on their mechanisms of action using cell and animal models.
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C. Polymers and their complexes:
Synthesis and structural studies of metal complexes with biologically relevant molecules using NMR spectroscopy, DFT computational methods and luminescence studies, with reference to their potential ability to chelate metals in vivo. Related studies on the interaction of tri- and divalent metal ions, such as aluminium(III), gallium(III), zinc(II) and copper(II) with hydroxyaromatic ligands, such as 8-hydroxyquinoline-5-sulfonate and with anionic synthetic polyelectrolytes using multinuclear NMR coupled with various other techniques, with the objective of the development of luminescent sensors for the analytical determination of these metals at low concentrations. Extension of these studies to biopolymers, such as nucleic acids, and other small molecules of biological interest, and the relevance to their potential ability to chelate hazardous metals in vivo.

Study on the structure and interaction of conjugated polymers and polyelectrolytes, with particular reference to their applications in biosensors of nucleic acids, sugars, proteins and in optoelectronic devices.

Analysis of the electronic structure and aggregation behaviour of conjugated polymers, studied as models for DNA, using nuclear magnetic resonance spectroscopy (NMR), computational methods based on density functional theory (DFT), small angle neutron scattering (SANS), UV/vis absorption and fluorescence and other photophysical techniques.
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D. NMR studies of metabolic fluxes:
We study intermediary metabolism using 2H, 13C and 15N stable isotope tracers. Glucose, protein and lipid homeostasis are studied in human subjects, in animal models of human diseases, and in isolated perfused animal organs.
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Equipment:
Varian VNMR 600 System with 3 observe channels (V600)

Probes:
5-mm PFG triple resonance I.D. (optimal sample volume = 600-700 μl)
5-mm PFG Broadband (optimal sample volume = 600-700 μl)
3-mm PFG triple resonance I.D. (optimal sample volume = 180-200 μl)
3-mm PFG Broadband (optimal sample volume = 180-200 μl)
4-mm PFG nanoprobe (HR-MAS)

The Varian VNMRS 600 MHz NMR spectrometer is an equipment part of the Portuguese National Network of Nuclear Magnetic Resonance Spectroscopy (PTNMR) as type A spectrometer (i.e. high field spectrometer). As such 70% of its time of operation is dedicated in priority to the PTNMR Users (i.e. registered Users with a project approved by PTNMR referees). Its conditions of use are the ones of a PTNMR type A spectrometer.

Note on the use of 3-mm microprobes: The 3-mm probes offer a ~40% increase in sensitivity over their 5-mm counterparts provided that the molecule of interest is soluble in the smaller volume and that the signal linewidths are preserved. For many biological samples that contain salt, e.g. protein solutions or tissue extracts, the smaller volumes result in higher salt concentrations leading to longer pulse widths, less efficient 1H-broadband decoupling, and increased sample heating. This latter effect may result in a drifting lock signal particularly if the lock solvent is D2O.
Varian UNITY 500 System (U500)

Probes:
5-mm I.D. (optimal sample volume = 600-700 μl)
10-mm Broadband (optimal sample volume = 1ml)
5-mm Low frequency (optimal sample volume = 600-700 μl)
5-mm PFG Broadband (optimal sample volume = 600-700 μl)

The Varian Unity 500 MHz NMR spectrometer depends on the LabRMN-CNC. Its conditions of use and access are following the ones of a PTNMR type B spectrometer (i.e. low field NMR spectrometer).
Data Backup Service

The LabRMN-CNC does not currently provide archive or backup service: Users are strongly recommended to backup their data during their NMR session.
Guide for NMR access and use:

1) Location:

The LabRMN-CNC is located in the ground floor of the Faculty of Medicine at the University of Coimbra, Pólo I (bus stop: Universidade). The local opens directly into the courtyard of the Medicine Building aside the principal staff entrance into the courtyard, opposite to the entrance for vehicles.

Parking inside the courtyard is reserved for Faculty cardholders. There is no visitor car park. The street parking in and around the University fills up quickly after 8:30 AM. Please be aware that cars either parked improperly or in designated sites for any length of time will almost certainly be clamped.

There are regular bus services connecting the University with coach and train stations.

The LabRMN-CNC facility is open to every User from 9:30 AM to 6:00 PM, from Monday to Friday, with a pause between ~ 1:00 PM - 2:00 PM. Access to the lab during the night or the weekend is restraint only to LabRMN-CNC Users with personal required access cards (Medicine Faculty and LabRMN-CNC). If doors are closed, Users are pleased to ring at the entrance (right of the door).

2) Spectrometer availability:

Equipment of LabRMN-CNC is not free access. The use of both NMR spectrometers is limited to CNC Users registered in the LabRMN-CNC Users database and to PTNMR Users with appropriate approved PTNMR projects.

New Users planning to access to service provided by the LabRMN-CNC are pleased to contact the NMR Unit Manager first or the NMR Unit Coordinator to discuss about their projects and their needs. They are also directly informed about the specific operating of the NMR Unit and the occasional costs of the services. After the meeting, Users are registered in the LabRMN-CNC Users database as in the [CNC NMR] mailing list. They receive the NMR time request forms and are allowed to request NMR time.

The organization and NMR time allocation in the LabRMN-CNC agree with standards of the National Network for Nuclear Magnetic Resonance (PTNMR) and the Foundation for Science and Technology (FCT). Both NMR spectrometers are following PTNMR access rules. NMR time not allocated to PTNMR Users is shared between CNC Users (members and outsiders of LabRMN-CNC).

NMR Service:
Along of its own research projects the LabRMN-CNC aims to support work of scientists of the national academic community as business environment, from public or private institutions, by providing a regular weekly service (called NMR Service), usually performed during the Saturday. This Service provides NMR experiments (mono and multidimensional experiments acquisition, analysis) for a large panel of nuclei. This NMR Service is exclusively performed by LabRMN-CNC staff. Users interested by the NMR Service are please to contact first the NMR Unit Manager for more informations.

3) Scheduling:

NMR time schedules of both spectrometers are planned on a monthly basis, generally starting the first Monday of the month.

NMR schedule is usually planned the Thursday before the coming month. Users wanting to request some NMR time must fully fill a PDF NMR time request form, specific to the spectrometer they want to use. Filled NMR time request forms must be send to the Unit Manager (emeric@cnc.uc.pt) before the Wednesday 8:00 PM of the “planning week”. Collection of requests starts the Monday of the “planning week”. (Please consult the online calendar to check the dates)

A preview of the new month NMR time schedule will be available the Thursday evening on the LabRMN-CNC online calendar. Any request for modification will be processed during the following Friday and weekend if necessary.

Possible OPEN TIME (free NMR time not distributed) will be distributed on demand during the same period. All request for OPEN TIME must be done with a new fully filled NMR time request form.

The definitive NMR time schedule will be posted on the online calendar and send by e-mail through the [CNC-NMR] mailing-list before the Sunday evening (usually the Friday evening).

Cancellations of NMR time must be immediately notified to the NMR Unit Manager, at least a day before. Non usage of time (without reasonable notification of cancellation) will result in a two weeks suspension.

4) Materials and Sample Preparation:

The LabRMN-CNC, as the PTNMR, does not provide any consumable. Users have to supply their own tubes and NMR solvents. For gHX Nano NMR probe Users, rotors from the LabRMN-CNC are available for use and have to be requested to the Unit Manager during NMR time booking.

For External Users: NMR samples may be prepared in the wet-lab next to the NMR room belonging to the LabRMN-CNC teams. The wet-lab is equipped with pipettes, a balance, a thermostated bath and micro-centrifuge. There is also a fridge / freezer to temporary store perishable samples. Please notice the wet-lab does not have fume hood or solvent evaporator.

Users are pleased to consider the following recommendations:
- use a deuterated solvent.
- use an intact and good quality NMR tube.
- avoid too big sticker on the tube (2 cm max glue on top of the tube) All samples must be labelled.
- use constantly the recommended amount of sample in the tube: 200 µL for 3 mm tubes and 650 µL for 5 mm tubes.
Pricing informations for the use of the LabNMR spectrometers (starting September 1st 2011)
I – Costs of the service for Academics (Non-Profit)/PTNMR Users
Spectrometer Spectral Acquisition (€ / hour)
With operator* Without operator**
VNMRS 600 10,00 7,00
Unity 500 8,00 5,50
* Experiments preparation time with operator charged.
** Cost for acquisition extra time after preparation.

Costs include the legal taxe IVA of 23%.
II – Costs of the services for companies (For-Profit)a
Spectrometer VNMRS 600 Unity 500
Cost (€) Cost (€)
1H 65,00 40,00
13C 190,00 120,00
31P 190,00 120,00
19F 190,00 120,00
13C-DEPT 190,00 120,00
COSY 190,00 120,00
HSQC 320,00 200,00
HMBC 570,00 360,00
NOESY 680,00 420,00
TOCSY 190,00 120,00
Package 1D* 390,00 240,00
Package 1D+2D** 1180,00 730,00
*includes 1H, 13C, 13C-DEPT
**includes 1H, 13C, 13C-DEPT, COSY, HSQC, HMBC

Costs include the legal taxe IVA of 23%.
a Costs are based on an average time of preparation and acquisition of the experiments. They include the use of the equipments as the preparation of sample and standard deutered solvants (CDCl3, D2O, DMSO, acetone).
Related with the sample quantity (MM), costs are valid for:
1H and 2D > 10 mg; 13C and 13C-DEPT > 200 mg; 31P > 10 mg; 19F > 10 mg
In case more acquisition time is needed than established for some experiments additionnal hours are charged (600 MHz 90€/h and 500 MHz 70€/h). Analysis of sample in relatively small quantity (concentration between 2 and 5 mg) is possible under conditions and a budget will be given after consultation of the service.
Processed spectra are send in electronic format (PDF) and acquisition files are available.
Elaboration of reports of spectra interpretations is possible. But due to projects specificities reports are charged in accordance with the time of their execution (110 € / hour).
III - Bio-NMR
Spectrum VNMRS 600
Cost (€)a
15N-HSQC 190,00
13C-HSQC 590,00
3D-HNCO 1490,00
3D-HN(CA)CO 1490,00
3D-HN(CO)CACB 1490,00
3D-HNCACB 1490,00
3D-15N-NOESY 2900,00
3D-15N-TOCSY 1490,00
3D-hCCH-TOCSY 2900,00
3D-HNHA 1490,00
3D 13C-NOESY 2900,00
Costs include the legal IVA tax of 23%.

a Costs are based on an average time of preparation and acquisition of the experiments for labelled samples (13C and 15N) with a standard concentration of 0.5 mM and an approximative molecular mass of 20 kDa.
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