Project Summary/Abstract: The Bruker timsTOF fleX includes a liquid chromatograph (LC), matrix assisted laser desorption ionization (MALDI-2) and electrospray ionization (ESI) sources, trapped ion mobility spectrometer (tims), and quadrupole- time-of-flight (QTOF) mass spectrometer combined into an instrument that can analyze metabolites, lipids, and proteins with high content or high throughput. Most importantly, the gap between these two experiments can be bridged easily when both are conducted on the same instrument; for example, metabolomics data can be effectively integrated with mass spectrometry imaging. To describe the individual components, the liquid chromatograph provides separation of analytes to increase depth of proteome and metabolome sampling. The two ion sources enable analysis of liquid samples (ESI) or arrays of samples co-crystallized with matrix (MALDI). The trapped ion mobility spectrometer (tims) provides data for collision cross section (CCS) calculations to further define each analyte as well as gas phase enrichment of molecules into packets with similar CCS-to-charge ratios for mass analysis. The QTOF provides mass spectrometry and tandem mass spectrometry with high resolution and accurate mass measurement. Vendor software provides support for data analysis in both metabolomics and proteomics as well as visualization and analysis of mass spectrometry imaging. The addition of this instrument to our Proteomics & Metabolomics Core (P&MC) complements our existing LC-MS resources, provides novel capability for ion mobility-mass spectrometry, replaces a MALDI instrument that was decommissioned last year, and enhances resources available for mass spectrometry imaging. As tumor heterogeneity, the tumor-immune interface, and spatial distribution of analytes in tumors now have increasing importance, existing analytical platforms must be modified to meet these needs and additional investments in instrumentation are needed. The Bruker timsTOF fleX complements existing LC-MS instruments and optimizes the value of investment in new technology by lowering sample input requirements, creating capability for spatial omics of tissues, supporting high throughput screening with MALDI-tims-MS, and ion mobility separation of isomeric lipids, metabolites, and post-translationally modified or chemically labeled peptides. Examples provided by Bruker have been confirmed by demonstration data from our own samples. This device also can be used to improve biomarker discovery and integrate molecular measurements with traditional pathology workflows. The placement in a core facility with highly trained staff and extensive institutional support will maximize both the availability to the user group and their benefit from access to the instrument. Together, these advantages are expected to significantly advance cancer metabolomics and proteomics applications at Moffitt Cancer Center to support existing and planned NIH/NCI-funded research and fuel the deve...