Our previous work 29 involved resonant excitation. This relies on high-energy, ultraviolet light (often provided by a 405 nm laser), which is detrimental to biological systems. Additionally, one-photon FLIM and PLIM techniques are known to suffer from high absorption and autofluorescence. If dynamic processes are to be studied in vivo as potential bioremediation strategies these drawbacks must be overcome by developing non-destructive analytical and imaging techniques. One such alternative approach is non-resonant, or multiphoton, excitation. The most common example of multiphoton excitation is two-photon excitation (2PE), which involves the simultaneous absorption of two photons via a transitory virtual state. Relaxation back to the ground state may then occur, releasing a single photon of higher energy than the incident radiation. As excitation relies on red or near-infrared (NIR) light it is less damaging to biological tissue, has deeper penetration depths and can result in lower autofluorescence. Additionally, the localised excitation volume allows control of excitation in the z-dimension, an advantage for microscopic investigations of spatially-complex samples. These benefits have led to the development and application of dyes for use in two-photon microscopy, 34 aimed at imaging particular cellular structures or tagging biomolecules of interest but little work has been carried out directly imaging an analyte of interest. To date, the only studies that have been carried out on the two-photon excitation of U VI O2 2+ have used crystalline samples, as well as cryogenic (liquid helium) conditions, or rather high-energy excitation wavelengths (532 nm). Here we demonstrate, for the first time, that uranyl(VI) emission can be induced by 2PE in the near infra-red, at room temperature and in amorphous phases. We then image uranyl(VI) with 2PE microscopy in yeast, shedding new light on the biosorption process, and in bacterial systems, allowing direct visualisation of the redox chemistry on the cell surface.