, Volume 397, Issue 8, pp 3409-3419
Date: 16 Jul 2010

Introduction of a 20 kHz Nd:YVO4 laser into a hybrid quadrupole time-of-flight mass spectrometer for MALDI-MS imaging

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A commercial hybrid quadrupole time–of–flight mass spectrometer has been modified for high-speed matrix-assisted laser desorption ionisation (MALDI) imaging using a short-pulse optical technology Nd:YVO4 laser. The laser operating in frequency-tripled mode (λ = 355 nm) is capable of delivering 1.5-ns pulses of energy at up to 8 μJ at 5–10 kHz and 3 μJ at 20 kHz. Experiments to improve beam homogeneity and reduce laser speckle by mechanical vibration of the fibre-optic laser delivery system are reported along with data from trial and tissue imaging experiments using the modified instrument. The laser appeared to yield best results for MALDI-MS imaging experiments when operating at repetition rates 5–10 kHz. Combining this with raster imaging allowed images of rat brain sections to be recorded in 37 min. Similarly, images of the distribution of peptides in “on-tissue” digest experiments from tumour tissues were recorded in 1 h and 30 min rather than the 8-h acquisition time previously used. A brief investigation of targeted protein analysis/imaging by multiple reaction monitoring experiments “on-tissue” is reported. A total of 26 transitions were recorded over a 3-s cycle time and images of abundant proteins were successfully recorded.