Protocol for Soft X-Ray Tomography

A paper describing a step-by-step sample preparation, data collection and cell segmentation protocol for full-rotation Soft X-ray Tomography (SXT)

This paper from the Advanced Light Source, LBNL, presents a detailed step by step protocol for full rotation soft X-ray tomography (SXT) of single cells.

The specimens are held in cylindrical sample holders which can be rotated through 360°, which eliminates artifacts caused by limited tilt tomography of flat sample holders.

SXT of single cells allows the visualisation of whole-cell morphology, including the nucleus and chromatin distribution, mitochondria, lipid droplets, endoplasmic reticulum, endosomes, lysosomes, and intracellular vesicles.

This provides information about organelle spatial distribution and changes in the cell ultrastructure organization caused by a viral infection or other conditions, as well as by the alteration in the biochemical composition. The acquisition of multiple tomograms from the same cell type and cell condition allows the microscopist to map and quantify the mutual arrangement of the subcellular components, thus providing an average cellular phenotype.


A protocol for full-rotation soft X-ray tomography of single cells

Jian-Hua Chen, Bieke Vanslembrouck, Valentina Loconte, Axel Ekman, Venera Weinhardt,