Improvements in fast mass microscopy for large-area samples

Mass spectrometry imaging (MSI) is a technique that analyzes chemical information and spatial distribution of surface analytes. Most MSI studies are conducted in microprobe-mode, in which a mass spectrum is collected for each pixel to create a mass image. Thus, spatial resolution, sample imaging area, and imaging speed are linked. In this mode, halving the pixel size quadruples the analytical time, which presents a practical limit on high spatial resolution MSI throughput. Fast mass microscopy (FMM) is, in contrast, a microscope-mode MSI technique which decouples spatial resolution and imaging speed. FMM circumvents the linear-quadratic relationship of pixel size and analytical time which enables increased the imaging size area and the analytical speed achievable. In this study, we implement instrument modifications to the FMM system including the addition of linear encoders that enable roughly 8.5× faster imaging than was previously achieved, allowing a 42.5 × 26 mm2 sample area to be imaged at a 1 µm pixel size in < 4.5 min. Linear encoders also enable the alignment of multi-pass images that increase image homogeneity and chemical signal. The applicability of FMM to large area samples has made it important to define the tolerance to height variations of the technique, which was determined to be at least 218 µm.