Axel Ekman, Jian-Hua Chen, Bieke Vanslembrouck, Valentina Loconte, Carolyn A. Larabell, Mark A. Le Gros, Venera Weinhardt
Soft X-ray tomography offers rapid imaging of whole, single cells with a few tens of nanometers spatial resolution without fixation or labeling. Herein, this technique is limited to specimens about 10 μm thick, such that applications of soft X-ray tomography of large human cells or multicellular specimens are not possible. A theoretical and experimental framework for soft X-ray tomography that enables extension of imaging volumes to 18 μm-thick specimens is developed. This approach, based on long depth of field and half-acquisition tomography, is easily applicable to microscopes equipped with a full-rotation specimen stage. This opens opportunities for imaging large human cells, such as those commonly seen in cancer research, as well as cell-to-cell interactions, where two or more cells occupy the same imaging volume.