Direct whole-mount imaging of fungal spores by energy-filtering transmission electron microscopy

Abstract

Whole-mount fungal spores were examined by energy-filtering transmission electron microscopy. Conidia of Penicillium species and Ustilaginoidea virens were suspended in distilled water and directly placed on a glow-discharged formvar-coated copper grid. Energy-filtered images were taken from 0 to 100 eV loss regions. Due to their considerable inherent thickness, their globose morphology was evident. In zero-loss images, the fungal spores appeared to have higher contrast in general, showing darker periphery than unfiltered images. Most spores in zero-loss images exhibited almost homogeneous electron density across the spores. The contrast was partially inversed in low-loss images where more details of the outer cell wall ornamentations of spores could be discerned than zero-loss images. As obvious advantages of whole-mount spore imaging, it allows for ensuring two-dimensional images with higher spatial resolution than light microscopy and conventional scanning electron microscopy. If a higher resolution is needed to observe fungal surface structures such as fimbriae and rodlet layers, or discriminate an outer sheath enveloping spores, whole-mount spore imaging can be employed to unravel structural details.