Capabilities and how to use the Velos Pro linear ion trap.
Slide 1
Welcome. This brief tutorial describes OHSU researchers using the Thermo Velos mass spectrometer in the OHSU Proteomics Shared Resource (PSR) core. This instrument is used on a walk-up/reservation basis and is operated by the researcher with minimal assistance by PSR core staff.
Slide 2
The Velos Pro Linear Ion Trap mass spectrometer is available for use by any OHSU researcher who has completed training from the OHSU PSR core staff.
Slide 3
The OHSU PSR core has high-resolution instruments for shotgun identification and quantification proteomics experiments. The linear trap is available for basic mass spectrometry measurements to support research such as expression and purification of protein constructs. Some of the more common uses are peptide mass measurements (less than 5000 Da) and smaller protein whole masses (5K to 50K Da). A variety of dissociation methods are available to fragment and sequence peptides, too. Reverse-phase liquid chromatography using C18 (peptides) or C4 (proteins) columns are available for purification/separations.
Slide 4
There are several options to introduce samples into the instrument. Direct infusion via syringe is the fastest way to obtain mass spectrometry data for your samples. If you have sufficiently purified starting material, HPLC or additional sample cleanup may not be needed. The samples must be in solutions that are compatible with mass spectrometry. Sample volumes and concentrations need to be within specified ranges to prevent overloading the system.
Slide 5
Care must be taken to work with compatible solvents for mass spectrometry. This slide contains information about approved solvents for direct infusion. The main concerns are that particulates and detergents, although common in regular sample handling, are not compatible with the instrument. The buffers also need some source of hydrogen ions (H+ in acidic solutions) to create ions that the mass spectrometer manipulates and measure.
Slide 6
HPLC chromatography with short reverse phase C18 or C4 columns are available for separations of analytes of interest. It may not be practical to start with very highly purified samples, and HPLC may be used to produce cleaner data for precise measurements.
Slide 7
After you are trained and qualified to operate the Velos Pro instrument, it is available on a first come first served basis. The instrument can also be reserved for planned experiments.
Slide 8
Operation of the instrument requires training by the OHSU PSR core staff. Analysis of the generated data is also part of the instrument use. The OHSU PSR core has proprietary vendor software on core computers available for use in room MRBA528. The PSR core staff provide training in the software use. The analyses of the types of data generated by the Velos Pro is within the capabilities of trained users, and users are expected to perform their own analyses. The core staff is happy to answer any questions and provide help.
Thank you!