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![[Figure 1]](gm5116fig1mag.jpg)
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Figure 1
Schematic workflow of the process envisioned for correlative hard X-ray nano-tomography and cryo-EM. The purpose of this workflow is to identify a region of interest (for example a gap-junction plaque at the cell–cell interface, illustrated here). To achieve this, (a) cells are subjected to high-pressure freezing within copper tubes (inner diameter 350 µm) or tissues are embedded in copper planchettes with a 200 µm cavity. These samples are then precisely trimmed using a diamond knife to yield a rectangular section measuring approximately 70 × 70 µm in area and 30 µm in thickness, optimized for hard X-ray nano-tomography. (b) An X-ray tomogram of the trimmed volume is acquired, allowing the identification of regions of interest (ROIs). (c) Thin sample layers are then extracted from selected ROIs using either cryo-electron microscopy of vitreous sections (CEMOVIS) or focused ion-beam (FIB) milling (*optionally guided by fluorescence light microscopy if fluorescence markers are present) for downstream cryo-EM analysis. |
![[Figure 1]](gm5116fig1.jpg)
 | STRUCTURAL BIOLOGY |
ISSN: 2059-7983
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