Instruments

Thermo Scientific Q Exactive UHMR
A benchtop Ultra High Mass Range (UHMR) hybrid quadrupole-Orbitrap instrument which enables analysis of extra-large molecules (m/z 350-80,000) with high mass accuracy. The UHMR is equipped with a higher-energy collision-induced dissociation (HCD) cell capable of acceleration voltages up to 300 V, as well as in-source trapping capabilities for improved declustering of ions. The Wysocki lab has two of these instruments and they are both modified with custom surface-induced dissociation (SID) devices. The UHMRs are primarily used for analysis of proteins and protein complexes by both direct infusion and online non-denaturing separation techniques. One instrument was purchased with funds from P41GM128577 and the other is part of a direct collaboration with Thermo Scientific.

Waters Synapt G2 HDMS
A quadrupole/travelling wave ion mobility (TWIM)/time-of-flight mass spectrometer. Ion mobility enables gas-phase ions to be separated by their size, shape, and charge. This affords a dimension of separation orthogonal to the standard m/z analysis. The instrument has been modified with a custom surface-induced dissociation (SID) device which can be placed before the IM region (SID-IM), after the IM region (IM-SID), or on either side of the IM cell to allow for SID-IM-SID. In addition, a linear field ion mobility cell can be installed and used to make direct collision cross section measurements without the need for calibration, as is necessary for TWIMS measurements. This instrument is mainly used for studies of large protein complexes.

Waters Synapt G2S HDMS
A newer generation of Waters Synapt G2 instrument with improved “step-wave” design after the ion source for improved sensitivity. Like the G2 this instrument has been modified with a custom surface-induced dissociation (SID) device that can be placed in the same possible configurations. This instrument is equipped with electron transfer dissociation (ETD) and has also been modified to incorporate a 193 nm laser for ultraviolet photo dissociation (UVPD). It is mainly used for protein complexes and more specifically for coupling SID and UVPD to do complex-down and top-down studies.

Thermo Scientific Exactive Plus EMR
A benchtop Orbitrap instrument with extended mass range (EMR) which enables analysis of large molecules (m/z 350-20,000) with high mass accuracy and is equipped with a variable pressure higher-energy collision-induced dissociation (HCD) cell. This instrument has been modified to include a high-mass range quadrupole mass filter and a custom surface-induced dissociation (SID) device. The EMR is primarily used for analysis of proteins and protein complexes by both direct infusion and online non-denaturing separation techniques.

Waters Synapt “G1” HDMS
The first generation Waters Q-TWIM-TOF platform. The high sensitivity of this instrument makes it particularly well suited to the study of low concentration samples. This instrument is primarily used for ion mobility studies of low concentration samples. A surface-induced dissociation (SID) device will be designed to replace the trap cell in this instrument.

Thermo Orbitrap Elite This hybrid instrument consists of a dual linear ion trap, an HCD cell and a high-field orbitrap analyzer. The orbitrap provides high resolution at up to 240,000. The dual linear ion trap allows faster scan rates. For tagged samples, the HCD cell allows lower m/z scan ranges. This instrument is also equipped with ETD which allows for fragmentation of longer peptides and top-down proteomics studies. The instrument is coupled to a 2D Waters nanoACQUITY which allows LC/LC-MS/MS analysis. It is suitable for many -omics studies.

Thermo LTQ Velos Pro This dual pressure linear ion trap allows fast scan rates. Along with trap HCD and ETD capabilities, this instrument is ideal for routine complex proteomic analysis. The instrument is currently coupled with a 1D Waters nanoACQUITY UPLC system for LC-MS/MS.