Correlative Light and Soft X-ray Microscopy in the Laboratory

Session

Poster Session

Authors

Paul Sheridan (1), Sergey Kapishnikov (1), Kenneth Fahy (1), William Fyans (1), Fergal O'Reilly (2, 1), Martina Donnellan (1), Tony McEnroe (1)

Affiliations

1. SiriusXT
2. UCD Dublin

Keywords

cryo soft X-ray tomography,  cryo fluorescence microscopy, correlative light and soft x-ray microscopy, 3D cell imaging, laboratory-scale device

Abstract text

Cryo-soft X-ray tomography is the unique technology that can image whole intact cells in 3D under normal and pathological conditions without labelling or fixation, at high throughput and spatial resolution [1-4]. The sample preparation is relatively straightforward; requiring just fast freezing of the specimen before transfer to the microscope for imaging. The technique can be correlated with cryo fluorescence microscopy to localize fluorescent proteins to organelles within the whole cell volume. Cryo-correlated light and soft X-ray tomography is particularly useful for the study of gross morphological changes brought about by disease or drugs.  For example, viral fluorescent tags can be co-localized to sites of viral replication in the soft X-ray volume.  In general this approach is extremely useful in the study of complex 3D organelle structure, nanoparticle uptake or in the detection of rare events in the context of whole cell structure. We will present data from our recently developed laboratory-scale SXT microscope which is propelling this unique and powerful workflow from the world of synchrotron research into the laboratory for everyday access [5,6]. Furthermore, the availability of cryo-soft X-ray tomography in the laboratory will accelerate the development of new light, electron and x-ray microscopy correlative and multimodal workflows [7]. An overview of the microscope and associated workflows will be presented along with imaging data from a range of case studies. 

References

[1] M Harkiolaki et al., Emerging Topics in Life Science 2 (2018), p. 18. doi: 10.1042/ETLS20170086

[2] J Guo and CA Larabell, Current Opinion in Structural Biology 58 (2019), p. 324. doi: 10.1016/J.SBI.2019.06.012

[3] G Schneider et al., Nature Methods 7 (2010), p. 985. doi: 10.1038/NMETH.1533

[4] FJ Chichón et al., Journal of Structural Biology 177 (2012), p. 202. doi:10.1016/J.JSB.2011.12.001

[5] Fahy et. al, (2024) Laboratory-based correlative cryo-soft X-ray tomography and cryo-fluorescence microscopy. In T. Müller-Reichert, & P. Verkade (Eds.), Methods in Cell Biology: Correlative Light and Electron Microscopy (vol. 187). Elsevier

[6] Fahy et. al., (2023) A Laboratory Based Soft X-ray Microscope for 3D Imaging of Whole Cells. Microscopy and Microanalysis 29, 1171-1172 https://doi.org/10.1093/micmic/ozad067.601

[7] https://clexm.eu/