The status of micro‐PIXE applications in investigations of ecophysiological aspects of the hyperaccumulation phenomenon in plants is reviewed. Measurements of elemental concentrations and distribution in organs, tissues and cells of plants hyperaccumulating Ni, Co, As, Mn, Zn and Cd, show that in most cases hyperaccumulated metals are stored in physiologically inactive tissues. However, some exceptions indicate that different physiological mechanisms are involved in metal transport, storage locations and detoxification. The influence of mycorrhiza on elemental concentration and distribution in the roots of the Ni‐hyperaccumulator Berkheya coddii is also discussed. Several herbivorous insects have been found feeding exclusively on Ni‐hyperaccumulating plants. A summary of micro‐PIXE contribution to studies aimed at explanation of their ability to survive in such extreme conditions is given. Copyright © 2011 John Wiley & Sons, Ltd.