Localisation of trace metals in metal-accumulating plants using μ-PIXE

Particle induced x-ray emission (PIXE) is a very sensitive technique that can quickly and reliably measure a wide range of elements simultaneously with high sensitivity. Using a focused microbeam, elemental distributions can be mapped with high spatial resolution. We demonstrate high-resolution mapping of metals in plant leaves at 5 mu m resolution and its application in detecting sites of metal accumulation in metal-accumulating plant tissues. The importance of biological sample preparation is discussed by direct comparison of freeze-substitution and freeze-drying techniques routinely used in biological sciences. The advantages and limitations of quantitative elemental imaging using these techniques are also discussed. Copyright (C) 2008 John Wiley & Sons, Ltd.