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An appraisal of ultramicrotomy, FIBSEM and cryogenic FIBSEM techniques for the sectioning of biological cells on titanium substrates for TEM investigation
Author(s) -
EDWARDS H.K.,
FAY M.W.,
ANDERSON S.I.,
SCOTCHFORD C.A.,
GRANT D.M.,
BROWN P.D.
Publication year - 2009
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.2009.03152.x
Subject(s) - materials science , transmission electron microscopy , foil method , titanium , focused ion beam , scanning electron microscope , sample preparation , microtome , biomedical engineering , nanotechnology , composite material , chemistry , optics , ion , chromatography , metallurgy , medicine , physics , organic chemistry
Summary 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.