Morphology visualization of irregular shape bacteria by electron holography and tomography.

In the current work, irregular morphology of Staphylococcus aureus bacteria has been visualized by phase retrieval employing off-axis electron holography (EH) and 3D reconstruction electron tomography using high-angle annular dark field scanning transmission electron microscopy (HAADF-STEM). Bacteria interacting with gold nanoparticles (AuNP) acquired a shrunken or irregular shape due to air dehydration processing. STEM imaging shows the attachment of AuNP on the surface of cells and suggests an irregular 3D morphology of the specimen. The phase reconstruction demonstrates that off-axis electron holography can reveal with a single hologram the morphology of the specimen and the distribution of the functionalized AuNPs. In addition, EH reduces significantly the acquisition time and the cumulative radiation damage (in three orders of magnitude) over biological samples in comparison with multiple tilted electron expositions intrinsic to electron tomography, as well as the processing time and the reconstruction artifacts that may arise during tomogram reconstruction.