MALDI Bruker Autoflex instrument use

WARNINGS:

Do not run metal or metal like compounds such as carbon nanotubes on the instrument, as this will destroy the detector and cause months of down time while the instrument is repaired as happened in the summer of 2020.
Do not adjust the Detector Gain without talking with the staff first. The parameter is under the mass range in the Detection tab so be careful not to accidentally change it.
If you have not used the MALDI since before 9/23, then use the default methods dated 9/23/20 to build or rebuild your method as this has been calibrated with the new detector.

BEFORE USING THE INSTRUMENT:

Watch this video to learn about MALDI sample preparation and spotting.

Reserve time using your FBS account, MSSelf Voyager MALDI Usage. Hours of instrument usage will be billed. <= 1hour is 1 hour, 1-2 hours is billed 2 hours, etc.

MALDI lab is MBB 1.420, normally open between 8am-5pm M-F. For after hours access, your PI will need to send a request for MBB building access and MBB 1.420 access to Maria Person at mperson@austin.utexas.edu

IN THE LAB:

Write your name/lab and plate/spots used into the black MALDI log book.
If you spot the calibration mix, note the number of the cal mix spot, type of cal mix and matrix, in the log book.
The data should be in the C:\Dat\Year_Month folder, e.g. 2016_Mar. You can create subfolders for your lab. Only these folders will be backed up monthly in case of computer crash.

CHCA, DHB and sinapinic acid are in the freezer in Eppendorf tubes in labeled 50 ml Falcon tubes. CM1 (1-3 kDa) and CM2 (2-5 kDa) made up in CHCA are in the CHCA tube and CM3 (5-20 kDa) are in the Sinapinic acid tube.
If you have problems talk to staff or email pmaf@austin.utexas.edu after hours. Note problems or strange behavior in the MALDI log book.

Warm up the instrument by turning on high voltage before spotting samples. The calibration will be stable after ~20 minutes.
Samples can be dried quicker by using the heat gun on cold or hot. Be careful on hot setting, the heat will melt plastic and plate will be dangerously hot.
Do not leave the plate holder in Eject mode and stay away from the ejected arm as bumping the arm will require a service call to fix. After removing a plate, return the plate holder to the instrument.

MALDI standard methods:

angiotensin–1000 Da, ACTH--2500 Da, Insulin–6 kDa, myoglobin–20kDa, 40 kDa, BSA–70 kDa, and IgG–100 kDa are all positive methods. Angiotensin, ACTH, and insulin are used with linear or reflector mode methods, other methods are only in linear mode. peptide_negative is negative method, all others are positive methods.

Chemistry MS Facility has a set of polymers that can provide a wide mass range calibration mix.

CLEANING PLATES:

Clean plates with soap and water, scrubbing with a Kimwipe in a circular motion to loosen debri, then with an Methanol and/or Ethanol 50:50 Water, then pure Methanol and/or Ethanol, scrubbing each time with the Kimwipe. Final rinse of plate with organic solvent. Allow to air dry tilted on side.