Journal
JOURNAL OF MICROSCOPY
Volume 234, Issue 1, Pages 16-25Publisher
WILEY
DOI: 10.1111/j.1365-2818.2009.03152.x
Keywords
Biomaterial; cryogenic FIBSEM; FIBSEM; human osteoblast; mesenchymal stem cell; titanium
Categories
Funding
- EPSRC [EP/E015379/1.]
- Engineering and Physical Sciences Research Council [EP/E015379/1] Funding Source: researchfish
- EPSRC [EP/E015379/1] Funding Source: UKRI
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Ultramicrotomy, focused ion beam scanning electron microscopy (FIBSEM) and cryogenic FIBSEM (cryo-FIBSEM) techniques, as developed for the controlled cross-sectioning of mesenchymal stem cells (MSCs) and human osteoblasts (HObs) on titanium (Ti) substrates for transmission electron microscopy (TEM) investigation, are compared. Conventional ultramicrotomy has been used to section cells on Ti-foil substrates embedded in resin, but significant problems with cell detachment using this technique restricted its general applicability. Conventional FIBSEM 'lift-out' procedures were found to be effective for the preparation of uniform sections of fixed and dehydrated cell/Ti specimens, but the control of cell staining remains an issue. Cryo-FIBSEM procedures used with an 'H-bar' sample geometry enabled the sectioning of fixed and hydrated cell/Ti specimens, but issues remain over ion beam-induced artefacts and control of frost on the sample foils.
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