The Center is establishing an advanced instrumentation facility that houses both high resolution NMR spectrometers and high resolution mass spectrometers that are directed by the senior members of the Center, and with full time manager/spectroscopists to ensure smooth operations and instrument maintenance.
*Support instrumentation for quality control and cell analysis (microscopy) is also available.*
The Thermo Tribrid Fusion is a Makarov-trap ("Orbitrap") Fourier-transform Mass Spectrometer (FTMS) capable of resolving power in excess of 350,000 @ m/z400, which was until recently achieved commercially only by ion-cyclotron resonance (ICR) MS. The Fusion instrument is capable of simultaneous, multi-mode ion processing and measurement via two collision/accumulator cells and two mass spectrometers (an Orbitrap and Ion Trap) with the ability to shuttle ions from either collision/accumulator cells to either MS. The strength of the Fusion is the ability to conduct very complex multistage tandem MS experiments, at great speed and extremely high resolution. This particular model is fitted with electron transfer dissociation (ETD) and internal lock-mass capabilities for ultra-stable mass mass reporting. The Fusion is typically combined with the Dionex UPLC capillary column electrospray or Advion Nanomate direct nanoelectrospray sample/ion sources.
- One instrument is primarily used for direct infusion, especially for lipid analyses. A second Thermo Tribrid Fusion FTMS Orbitrap is interfaced to chromatography, in particular the Dionex ion chromatography system, which can also interface to the Agilent triple quadrupole ICP-MS (below).
The Bruker Daltonics SolariX XR is an ultra-high resolution, accurate-mass ion cyclotron resonance mass spectrometer (ICR-MS). Incorporating the new ICR Paracell with absorption mode processing, it is capable of resolving power up to m/∆m = 10 million. In practice this means the SolariX XR can determine unambiguous molecular formulae (m.f.) directly from MS1 by interpreting the natural abundance isotopic fine structure; the advantage of being able to do this with just MS1 is that nanoelectrospray infusion can be used for m.f. identification, achieving both higher speed with higher signal-to-noise, while conducting molecular formula determinations on thousands of metabolites at once. The SolariX is also used to confirm unambiguously the presence or lack of interferences in samples destined to run on the Fusion FT-MS, itself a high-resolution mass spectrometer. As with the Fusion, the SolariX is also capable of multistage tandem MS experiments in combinations with Dionex UPLC capillary column electrospray or Advion Nanomate direct infusion nanoelectrospray, and adds medium vacuum MALDI capability with upgrade options to MALDI-imaging.
The Thermo ITQ 1100 ion trap gas chromatograph-mass spectrometer (GC-MS) analyzes low molecular weight polar derivatized metabolites, which has been a mainstay in our metabolic work since the early 1980's. The ITQ is capable of multistage tandem MS experiments, enhancing its selective trace quantification capability. There is also a Thermo Polaris Q GC-MS, also capable of multistage tandem MS experiments, that is dedicated to work on volatile metabolites without derivatization.
One system will be dedicated to gas analyses for determining C-13 enrichment in CO2.
An Agilent triple quad ICP MS was installed January 2015. This is an ultra high sensitivity mass spectrometer with high elemental ion selectivity across the periodic table. A Dionex ion chromatography module that resolves negatively charged analytes was installed Fall 2015. This enables speciation of compounds detected by element.
The NMR instrumentation currently comprises one high field Agilent spectrometer operating at 14.1 T with 4 channels, and a 3 mm HCN cold probe. The system has full automation via autotune and cooled autosamplers.
Two additional spectrometers were installed in 2016. A three-channel dual receiver wide bore 9.4 T microimaging/spectroscopy system for small animal and tissue-based imaging and spectroscopy, and a 16.45 T four channel dual receiver spectrometer equipped with two cryoprobes and cooled sample changer. Data stations running NMR analysis software are part of the Center.
14.1 T Spectrometer
The 14.1 T Agilent DD2 spectrometer is dedicated to metabolic studies. The instrument includes autotune for the probes, ensuring optimal pulse calibrations, with a 96-place random access cooled autosampler that can accept any tube size or configuration. The sample can be maintained at 5 °C to maintain integrity when larger numbers of samples are to analyzed. The instrument is equipped with two probes:
- a room temperature 5 mm OneProbe capable of any double resonance experiments with 1H or 19F observe
- a 3 mm cold HCN 13C-enhanced probe which is the workhorse metabolomics probe for proton-detected 13C or 15N SIRM experiments using small volume samples (< 100 µL with 3 mm sample tube; <50 µL with a 1.7 mm sample tube).
- Direct 13C observe on this probe is more sensitive than a broad band RT probe.
- Triple resonance with 2H decoupling is standard on the cold probe.
16.45 T Spectrometer
The Bruker Avance III spectrometer was installed September 2016. The system feature 4 channels, two receivers and a cooled SampleCase
sample changer and two autotune cryoprobes.
- 5 mm inverse quad probe (HCNP)
- 1.7 mm HCN 13C-enhanced probe for proton-detected 13C or 15N SIRM experiments using small volume samples (< 50 µL with 1.7 mm sample tube).
- Direct 13C observe on this probe is more sensitive than a broad band RT probe16.45 T Spectrometer
9.4 T Wide Bore Spectrometer
The Bruker Avance III spectrometer was installed December 2016. The system feature 3 channels, two receivers, microimaging with 120 G/cm a,x,z gradients and surface coils tuned to 31P and 13C
- 10 mm XH probe
Particle counting and sizing
A Malvern Instruments Nanosight300 has been installed (July 2016). This instrument counts particles in the range ca. 10-2000 nm in diameter, and also estimates the hydrodynamic radius from the translational diffusion coefficient, and the corresponding size distribution of suspended particles.
Fluorescence detection (excitation at 489 nm) via filters extends the capabilities to counting all particles present, and those labeled with a specific fluorescent tag.
An Olympus Fluoview FV1200 confocal/brightfield fluorescence microscopy system was installed Summer 2016. The system features 6 lasers, 4 objectives. DIC, scanning stage, spectral scanning, two GaAs PMTs, and an environmental chamber for live cell imaging. Both a regular (Microm HM355) and a cryomicrotome (Thermo HM 5252 NX) have been installed in CESB.