Metabolic Flux and Enrichment Studies using MIMOSA

As part of Yale Cancer Center (YCC) efforts to enhance membership access to metabolomics and metabolism technologies, the Kibbey laboratory will provide at cost steady state measurements of mitochondrial metabolism using the MIMOSA (Mass Isotopomer MultiOrdinate Spectral Analysis) platform (1). Consultative/collaborative design, application and interpretation of ¹³C labeled MIMOSA fluxomic measurements in cultured cells are key features of the service.

MIMOSA is a mass spectroscopy technique using stable isotopes to quantitatively measure the discrete rates of sequential individual reactions in glycolytic and mitochondrial metabolism. Such measurements can be used to identify differences in fuel usage (e.g., glycolysis vs. beta-oxidation), non-oxidative (reductive carboxylation, anaplerosis and cataplerosis) contributions to the TCA cycle. Additional development of the resource for the analysis of other metabolic fluxes may be possible.

Please Note: The typical turnaround time for data from these services is on average 2-3 months but can vary depending on laboratory work load and the complexity of the study.

Alves TC, Pongratz RL, Zhao X, Yarborough O, Sereda S, Shirihai O, Cline GW, Mason G, Kibbey R, (2015) Integrated, step-wise, mass-isotopomeric flux analysis of the TCA cycle. Cell Metab. 2015 Nov 3;22(5):936-47. doi: 10.1016/j.cmet.2015.08.021. Epub 2015 Sep 24. PMID: 26411341

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LC/MS/MS MIMOSA Study:

• Internal - $208/sample
• External - Please request quote

Sample cost includes LC/MS/MS sample plate preparation, plate run, peak integration and data processing/analysis/interpretation including isotopologue enrichment with appropriate phi calculations and metabolite concentration data.

Additional costs:

Full-service labeling study -

• Internal: $999
• External - Please request quote

The full-service labeling study is a one-time fee per study day including the stable isotopes [U13C-glucose, 1,2-13C-glutamine or U13C-lactate], the dynamic or steady state labeling experiment and cell quenching in preparation for the LC/MS/MS MIMOSA run. This service does not include but is in addition to the LC/MS/MS study run as a cost per sample described above.

For dynamic metabolic flux studies, it is strongly recommended that a pilot time course experiment be performed prior to the full LC/MS/MS study in order to determine sample sensitivity on the instrument and to determine metabolic and isotopic steady state for the particular cell samples. Please inquire for discounted pricing on the pilot experiment.

Method development-

• Internal: $770/day
• External: Please request quote

An additional fee for method development of new metabolites not in our MIMOSA central carbon metabolite panel will be applied if required. Generally, 3 new metabolite standards can be optimized per day. Standards are to be provided by the customer for this service. The price for this analysis is based on study assistance and instrument time needed for typical method development of 1-3 standards maximum.

Study assistance -

• Internal: $134/hour
• External - Please request quote

For additional sample preparation assistance that does not fall under the MIMOSA services described above is based on a per hour fee. Additional study assistance is required for samples other than cells submitted in the standard 6-well plate that require extra time-consuming steps such as rodent plasma/tissue processing from stable isotope label infusion studies or unlabeled tissues/plasma for metabolite concentrations.