CRI’s Metabolomics Facility is a shared resource located within the CRI as part of the Genetic and Metabolic Disease Program. The facility offers state-of-the-art metabolomics analysis and is available to scientists at CRI, UT Southwestern and collaborating institutes.
The Metabolomics Facility offers the following analytical methods:
- Basic metabolite quantitation for glucose, lactate, glutamate and glutamine.
- Measurements of respiration and extracellular acidification.
- Targeted metabolomic profiling (up to 180 metabolites).
- Stable isotope tracing/metabolic flux analysis.
If you would like access to the facility, please contact Lauren Zacharias.
Essentially every basic biological process, including energy production, growth, waste disposal and gene expression, requires proper control of metabolism. Normal tissue function cannot occur without precise control of metabolism, and as a result, many diseases involve perturbed metabolism at the cellular level. Such diseases include rare “inborn errors of metabolism” caused by inherited mutations in metabolic enzymes and common diseases like cancer, diabetes and heart disease.
Our facility uses an array of techniques to assess metabolism in living tissues. These techniques include:
- Target metabolite detection. These simple approaches quantify up to a small handful of metabolites (sometimes one metabolite) with a high degree of accuracy. An example is the measurement of lactate in tissue culture medium from glycolytic cancer cells.
- Targeted metabolomics. Targeted metabolomics analyzes a set of metabolites (ranging from a handful to a few hundred) simultaneously. For example, one could choose a set of 100 metabolites from informative positions within a metabolic network and then use mass spectrometry to determine the absolute or relative abundances of these molecules from a tissue sample, blood, culture medium, etc.
- Unbiased metabolomics. Unbiased metabolomics detects all metabolites, known and unknown, within a sample. This approach is less constrained by a priori knowledge than targeted metabolomics because it does not require the user to preselect a subset of metabolites to monitor.
- Isotope tracing/metabolic flux analysis. These techniques involve labeling with a stable isotope (e.g. 13C), complemented by computational analysis of isotope enrichment in metabolites downstream of the labeled nutrient, to understand flow through metabolic pathways.
CRI’s Metabolomics Facility provides services through a cost-sharing approach in which users are charged according to instrument time. Our staff consults with potential users to discuss experimental design and costs. Users then submit a sample form to estimate costs of the experiment. Staff is also available to discuss data interpretation after the experiment. To get started, please contact Lauren Zacharias.
- NOVA BioProfile 4 (measures glucose, lactate, glutamine and glutamate from tissue culture medium).
- Hansatech Oxygraph (measures oxygen consumption).
- Seahorse XF96 Extracellular Flux Analyzer (measures oxygen consumption and extracellular acidification).
- Hitachi L-8900 Amino Acid Analyzer.
- Gas chromatography/mass spectrometry (metabolite detection/isotope enrichment):
- GC/MS#1: Agilent 7890 networked to Agilent 5973 Mass Selective Detector.
- GC/MS#2 and #3: Agilent 7890 networked to Agilent 5975 Mass Selective Detector.
- AB SCIEX QTRAP 5500 LC/triple quadrupole mass spectrometer (targeted metabolomics/isotope enrichment).
- Agilent 6550 iFunnel LC/quadrupole-time of flight mass spectrometer (unbiased metabolite profiling).