Cell culture methods have been developed in efforts to produce biologically relevant systems for developmental and disease modeling, and appropriate analytical tools are essential. Knowledge of ultrastructural characteristics represents the basis to reveal in situ the cellular morphology, cell-cell interactions, organelle distribution, niches in which cells reside, and many more. The traditional method for 3D visualization of ultrastructural components, serial sectioning using transmission electron microscopy (TEM), is very labor-intensive due to contentious TEM slice preparation and subsequent image processing of the whole collection. In this chapter, we present serial block-face scanning electron microscopy, together with complex methodology for spheroid formation, contrasting of cellular compartments, image processing, and 3D visualization. The described technique is effective for detailed morphological analysis of stem cell spheroids, organoids, as well as organotypic cell cultures.
- Klíčová slova
- 3D visualization, Image reconstruction, Image segmentation, Morphology, Organoid, SBF-SEM, Scanning electron microscopy, Serial block-face, Spheroid, Stem cell, Ultrastructure,
- MeSH
- buněčné sféroidy ultrastruktura MeSH
- embryonální kmenové buňky ultrastruktura MeSH
- kultivované buňky MeSH
- lidé MeSH
- mikroskopie elektronová rastrovací metody MeSH
- počítačové zpracování obrazu MeSH
- zobrazování trojrozměrné metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
