Multiphoton and magnetic resonance imaging of Barley embryos: comparing micro-imaging techniques across scale and parameter barriers

Multiphoton stimulated autofluorescence microscopy and Magnetic resonance imaging (MRI) address different molecular properties of the sample and reach to a different length scale. MRI maps density or mobility of nuclei (here: hydrogen), and targets at whole objects from the scale of sub-millimetres to meters. Multiphoton imaging profits from the nonlinear absorption of light in the focus of a femtosecond laser source stimulating the autofluorescence of biomolecules. As this effect relies on a high light intensity the accessible field of view is limited, but the resolution is very high. Studying a plant embryo (barley) we compare the two techniques. At 770 nm excitation the cell walls of the embryo exhibited significant autofluorescence, allowing for a subcellular resolution. While details where imaged with an objective of N.A. 1.3, an overview was generated with a N.A. as low as 0.25. The overview image as well as merged images and tomographical data were used to link the high-resolution optical data with the three-dimensional highresolution MR images. There, images of the proton density were acquired using a standard 3D spin-echo imaging pulse sequence. While the optical high-resolution data provides a field of view restricted to only a small part of the embryo, the MR image contains the whole grain. Bridging the scales it might be possible to trace transport of e.g. nutrients from large structure of the plant to the cellular level.