This paper introduces multielement Z-tag X-ray fluorescence (MEZ-XRF), a novel high-speed, highly multiplexed bioimaging method. MEZ-XRF uses X-ray-sensitive reporter tags (Z-tags) based on different elements to achieve fluorescence-type imaging. Combined with signal amplification by exchange reaction (SABER), MEZ-XRF enables parallel imaging of numerous markers at subcellular resolution in various samples. The method is demonstrated to be nondestructive and capable of multiscale imaging, allowing for rapid tissue overviews followed by high-resolution scans of specific regions of interest. MEZ-XRF's unique capabilities are benchmarked against imaging mass cytometry (IMC) and showcase its potential for advanced multiomic tissue analyses.

Merrick Strotton, Tsuyoshi Hosogane, Marco di Michiel, Holger Moch, Zsuzsanna Varga, Bernd Bodenmiller