A combination of soft X-ray and laser light sources offer 3D high content information on the native state of the cellular environment

Abstract

Beamline B24 is a life sciences correlative cryo-imaging beamline at Diamond Light Source. B24 uses a combination of conventional and super-resolution visible-light fluorescence microscopy and soft X-ray tomography (cryoSXT) to provide 3D imaging of the cellular landscape at a resolution up to 25 nm in cryo-preserved biological samples up to 12 μm thick. B24 offers user-friendly, semi-automated 3D correlative cryo-imaging through an integrated platform of methods that encompass (a) sample preparation and evaluation, (b) data collection and processing and (c) data analysis and correlation. CryoSXT fills the current resolution gap between fluorescence and electron microscopy while cryo-structured illumination microscopy provides the additional dimension of chemical localization within the same cellular ultrastructure captured by cryoSXT. Beamline instruments can be accessed biannually by academics and industry globally through peer-reviewed standard and rapid access proposal processes. The B24 user base is primarily academic research groups studying cell function and cytopathology in biological systems ranging from viruses and algae to mammalian cells and proto-tissue complexes. Future work will consolidate development efforts and experiences gained thus far to enable high-throughput data collection. Special emphasis is placed on the delivery of other integrated advanced imaging methods such as X-ray absorption near-edge spectroscopy and phase contrast.