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September 5th, 2025
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Synchrotron Soleil
cell biology
bioRxiv

Bridging the resolution gap in cryo-CLEM by introducing cryo-SXT: cryo-CLXEM

Groen, J.Open in Google Scholar•Gazi, A.Open in Google Scholar•Kapishnikov, S.Open in Google Scholar•Brelot, A.Open in Google Scholar•Vos, M.Open in Google Scholar•Enninga, J.Open in Google Scholar•Pereiro, E.Open in Google Scholar•Sartori-Rupp, A.Open in Google Scholar

Cryo-imaging in cellular biology provides the means to visualize the cellular interieur at close-to-native conditions. A cornerstone in the field has been cryo-correlative light and electron microscopy (cryo-CLEM), with cryo-visible light fluorescent microscopy (cryo-VLFM) providing the specificity by tagging macromolecules or structures and cryo-electron tomography (cryo-ET) for the structural details at molecular level. The large resolution gap between these techniques, however, is limiting this correlative workflow as cryo-ET targets are often smaller than the resolution limit of cryo-VLFM. Here we introduce cryo-soft X-ray tomography (cryo-SXT) as an intermediate step that can compensate for the partial view and limited resolution of cryo-VLFM by providing invaluable cellular context information in 3D to the cryo-ET dataset within an integrated workflow.

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