Bacterial strains were isolated from the oral cavity of healthy volunteers and grown in the presence of Hg-ions (1-10 ppm) or arsenate ions at concentrations of 0.1-1.0%. To elucidate how bacteria take up and transform toxic metals inside the cells, we performed ion imaging and depth profiling with time-of-flight secondary ion mass spectrometry (ToF-SIMS). This analysis relies on the use of a pulsed ion beam to ionize surface molecules that can be extracted into a time-of-flight mass spectrometer. By combining the pulsed ion beam with another ion beam in direct current (DC) mode, depth profiles are obtained as a result of consecutive removal of surface layers. The spatial resolution of the depth profile along the y-axis is in the range of a few nanometres, and the spatial resolution along the x-and z-axes is in the lm range. The ToF-SIMS analysis was performed on crude biofilms of bacteria air-dried at aluminium foil surfaces, allowing subcellular resolution along the y-axis. The mercury ions were found transformed to methylmercury preferably in the periplasmic space, and the arsenate ions were found reduced to arsenite inside the cells, close to the cell membrane. The data are discussed in relation to current concepts in bacterial resistance to metals and antibiotics.Protection of human beings against toxic metals is a main issue of health care, especially in developing countries. Apart from direct toxic action of the metal, toxic metals are also suspected to induce resistance of bacteria to antibiotics [1,2], which is a threat to human health care. To find strategies of protection against toxic metals, there is a call for more knowledge on the effect of exposure and mechanisms of action.Imaging ToF-SIMS [3] is a method of analysis allowing localization of different chemical species of metals and organic molecules in cells and tissues. The spatial resolution makes subcellular localization of metal compounds possible, thus allowing analysis of the complete metabolism of metals in cells. This technique was adopted to study the fate of As-and Hg-species in oral bacteria that were found to be resistant when exposed to arsenate-or Hg-ions.In principle, the resistance of bacteria to toxic metals depends on one or combinations of the following subprocesses in the bacterial reaction to the metals: firstly, the influx of metals into the cells occurs via porins [4][5][6][7], which may be opened or closed by the contact with toxic metals and the number of pores may be down-regulated as a consequence of the porin-metal interaction [8]. Secondly, the metals are modified by enzymes inside the cells [9,10]. Oxidation, reduction and methylation of the metals are common results of bacterial enzyme action [10][11][12]. Finally, the efflux of the enzymemodified metals occurs via porins [7,13].The enzyme action on metals facilitates the efflux and is a key event in bacterial resistance to toxic metals. The enzymes that are active in processing metals in bacteria may either be intrinsic or acquired by infection of the cel...