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Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF MS) imaging of surfaces and tissues is a rapidly evolving technique having great potential in the field of biosciences. In earlier times, acquisition of a single high-resolution MS image could take days. Despite the recent introduction of high-repetition rate lasers to increase sample throughput of axial TOF MS instruments, obtaining a high-resolution image still requires a few hours. This paper shows that a substantial increase in the throughput of the TOF MS-based tissue imaging can be achieved by incorporating a mirror providing high-speed precision scanning of the laser beam along the sample surface. Equipped with the scanning mirror, a laboratory-built axial MALDI TOF MS instrument utilizing a 4-kHz UV laser recorded a 100 × 100 pixel MS image in ~11 min using 100 laser shots per pixel. This is almost an order of magnitude faster when compared to a modern commercial instrument equipped with 1-kHz laser.

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