Precise and economic FIB/SEM for CLEM: with 2 nm voxels through mitosis

Luckner, Manja and Wanner, Gerhard (2018): Precise and economic FIB/SEM for CLEM: with 2 nm voxels through mitosis. In: Histochemistry and Cell Biology, Vol. 150, No. 2: pp. 149-170

Abstract

A portfolio is presented documenting economic, high-resolution correlative focused ion beam scanning electron microscopy (FIB/SEM) in routine, comprising: (i) the use of custom-labeled slides and coverslips, (ii) embedding of cells in thin, or ultra-thin resin layers for correlative light and electron microscopy (CLEM) and (iii) the claim to reach the highest resolution possible with FIB/SEM in xyz. Regions of interest (ROIs) defined in light microscope (LM), can be relocated quickly and precisely in SEM. As proof of principle, HeLa cells were investigated in 3D context at all stages of the cell cycle, documenting ultrastructural changes during mitosis: nuclear envelope breakdown and reassembly, Golgi degradation and reconstitution and the formation of the midzone and midbody.

• Item Type: Journal article
• Faculties: Biology > Department Biology I
• Subjects: 500 Science > 570 Life sciences; biology
• ISSN: 0948-6143
• Language: English