title: Focused ion beam (FIB)/scanning electron microscopy (SEM) in tissue structural research.
abstract: The focused ion beam (FIB) and scanning electron microscope (SEM) are commonly used in material sciences for imaging and analysis of materials. Over the last decade, the combined FIB/SEM system has proven to be also applicable in the life sciences. We have examined the potential of the focused ion beam/scanning electron microscope system for the investigation of biological tissues of the model organism Porcellio scaber (Crustacea: Isopoda). Tissue from digestive glands was prepared as for conventional SEM or as for transmission electron microscopy (TEM). The samples were transferred into FIB/SEM for FIB milling and an imaging operation. FIB-milled regions were secondary electron imaged, back-scattered electron imaged, or energy dispersive X-ray (EDX) analyzed. Our results demonstrated that FIB/SEM enables simultaneous investigation of sample gross morphology, cell surface characteristics, and subsurface structures. The same FIB-exposed regions were analyzed by EDX to provide basic compositional data. When samples were prepared as for TEM, the information obtained with FIB/SEM is comparable, though at limited magnification, to that obtained from TEM. A combination of imaging, micro-manipulation, and compositional analysis appears of particular interest in the investigation of epithelial tissues, which are subjected to various endogenous and exogenous conditions affecting their structure and function. The FIB/SEM is a promising tool for an overall examination of epithelial tissue under normal, stressed, or pathological conditions.
authors.affiliation: Department of Biology, Biotechnical Faculty, University of Ljubljana, Vecna pot 111, SI-1000, Ljubljana, Slovenia.
authors.forename: Vladka
authors.initials: V
authors.lastname: Leser
authors.forename: Marziale
authors.initials: M
authors.lastname: Milani
authors.forename: Francesco
authors.initials: F
authors.lastname: Tatti
authors.forename: Ziva Pipan
authors.initials: ZP
authors.lastname: Tkalec
authors.forename: Jasna
authors.initials: J
authors.lastname: Strus
authors.forename: Damjana
authors.initials: D
authors.lastname: Drobne
mesh_terms: D000818:Animals; D004064:Digestive System / Q000648:ultrastructure; D006801:Humans; D033321:Isopoda / Q000648:ultrastructure*; D008855:Microscopy, Electron, Scanning / Q000379:methods*