X-ray lightsheet microscopy demonstrates nanometre-scale 3D marker positioning

Determining positions through thin X-ray sections

An international research group led by RIKEN researchers reported using X-ray lightsheet microscopy at SPring-8 to perform 3D localization from non-destructive measurement data while minimizing radiation-damage effects through short exposures. In a silica-sphere validation experiment, it reported 7 nm in-plane and 2 nm depth positioning precision for nanoparticle markers. The group also reported observing the 3D distribution of 10 nm markers inside hamster ovarian cells without destructive measurement in air.

The report describes conventional 3D localization super-resolution fluorescence microscopy as relying on controlled fluorescence blinking to keep nearby signals from overlapping, a process that can lengthen imaging time.

According to the research group, a thin X-ray lightsheet separated the excited marker signals spatially, allowing non-blinking scintillating nanoparticles to be used for short-exposure 3D super-resolution imaging.

The reported approach uses the geometry of a thin X-ray lightsheet rather than time-controlled blinking to separate fluorescent signals. The group says this could support non-destructive 3D observation of cell interiors and industrial products, but those future applications remain an expectation.