Cores for Vision Research Administration Ocular Microanatomy Core (OM Core) Visual Function and Non-invasive Ocular Imaging Core (VFNOI Core) Ocular Mass Spectrometry, Lipidomics, and Proteomics Core (OMSLP Core) Ophthalmic Testing Core (OTC) Ocular Mass Spectrometry, Lipidomics, and Proteomics Core (OMSLP Core) Home Research Cores For Vision Research Ocular Mass Spectrometry, Lipidomics, and Proteomics Core (OMSLP Core) Ocular Mass Spectrometry, Lipidomics, and Proteomics Core Cores of Vision Research within the University of California Irvine (UCI) will strengthen and enhance NEI R01-funded research. The project aims to support the major expansion of vision research currently underway at UCI. Our goal is to broaden and optimize research capabilities, support and expand existing research projects, and help establish new studies, ultimately boosting productivity and strengthening vision research at UCI. It is sponsored by the NEI P30 grant, P30EY034070-01. INITIAL SERVICE REQUEST FORM Rachel Martin, PhD Core Director, Ocular Mass Spectrometry, Lipidomics & Proteomics Fangyuan Gao, PhD Core Manager, Ocular Mass Spectrometry, Lipidomics & Proteomics Core Reservation sign-up link for Equipment use Equipment used by the Ocular Mass Spectroscopy, Lipidomics, and Proteomics Core is located on the second floor of the Gillespie building. This is where most of the labs forming the Center for Translational Vision Research (CTVR) are housed, which makes the location of the Core convenient and easily accessible. This space is also in close proximity to all vision researchers in the School of Medicine. The two mass spectrometers that are part of the MSLP Core are housed in separate rooms, one located in Dr. Skowronska-Krawczyk laboratory and the other one in Dr. Palczewski’s laboratory. Both are available for use by vision science users. Reservation sign-up link for equipment use Major Equipment OMSLP core The OMSLP core has two mass-spectroscopy instruments devoted solely to vision research. Thermo Scientific LXQ Linear Ion Trap Mass Spectrometer equipped with APCI and ES sources dedicated to analyses of small molecules, lipids and peptides. This instrument is currently maintained by Dr. Palczewski laboratory and is used by members of the CTVR laboratories. Thermo Scientific QExactive LC-MS system with APCI and ES sources and a Vanquish Binary Ultra High-Pressure Liquid Chromatograph. This newly-acquired instrument will be used to analyze untargeted and targeted lipidomics as well as identification of small molecules. This instrument was purchased with funds provides by several investigators within CTVR. This instrument is currently maintained by the Skowronska-Krawczyk laboratory and is used by members of the CTVR laboratories. LTQ XL™ linear ion trap mass spectrometer LTQ XL™ linear ion trap mass spectrometer can obtain high sensitivity full scan MS along with in-depth MSn (CRM) capabilities. A single quadrupole offers Selected Ion Monitoring. A triple quadrupole adds Selected Reaction Monitoring. A linear ion trap goes further by enabling Consecutive Reaction Monitoring where a selected ion is fragmented into a product ion and the resulting product ion further fragmented in additional consecutive steps. Each additional fragmentation step makes compound identification more certain and facilitates structural characterization. It offers high quality data with mass range 15 to 200 m/z; 50 – 2000 m/z; 200 – 4000 m/z. QExactive Mass Spectrometer QExactive mass spectrometer can be operated in several scan modes, including: full-scan MS (full MS), all-ion fragmentation (AIF) in the HCD collision cell, SIM for single ion detection, and parallel reaction monitoring (PRM) for detecting all fragment ions in parallel. Fragments generated in-source or in the multipole collision cell by AIF provide additional information that complements full-scan data. AIF fragments all ions in a single fragmentation event without precursor ion isolation and records all fragment ions in a single mixed spectrum. MS/MS (MS2) data can be collected using data-dependent acquisition (DDA) where precursor ions selected using predetermined rules are subjected to MS/MS, or by using data-independent acquisition (DIA) where a full scan is followed by a number of wide isolation MS2 scans, which together cover the same isolation range as the preceding full scan. The benefit of DIA is the ability to capture both all data and MS2 HR data scans without need for additional sample injections, enabling the acquisition of full information for all compounds and their fragments. The ability to perform fast polarity switching allows acquisition of both positive and negative scans within a single acquisition method, eliminating the need for extra sample injections, which is of particular importance when only limited amounts of sample are available. Positive and negative polarity switching can be performed on the chromatographic timescale and does not require instrument recalibration. The QExactive series of mass spectrometers provide up to six orders of magnitude of linear quantitative dynamic range and up to five orders of intra-scan dynamic range for accurate quantitation in low-level components in complex matrices. In addition, these mass spectrometers are easily calibrated in just a few minutes and maintain that calibration for days at a time.