Electron and ion imaging of gland cells using the FIB/SEM system

Abstract

SummaryThe FIB/SEM system was satisfactorily used for scanning ion (SIM) and scanning electron microscopy (SEM) of gland epithelial cells of a terrestrial isopod Porcellio scaber (Isopoda, Crustacea). The interior of cells was exposed by site-specific in situ focused ion beam (FIB) milling. Scanning ion (SI) imaging was an adequate substitution for scanning electron (SE) imaging when charging rendered SE imaging impossible. No significant differences in resolution between the SI and SE images were observed. The contrast on both the SI and SE images is a topographic. The consequences of SI imaging are, among others, introduction of Ga + ions on /into the samples and destruction of the imaged surface. These two characteristics of SI imaging can be used advantageously. Introduction of Ga + ions onto the specimen neutralizes the charge effect in the subsequent SE imaging. In addition, the destructive nature of SI imaging can be used as a tool for the gradual removal of the exposed layer of the imaged surface, uncovering the structures lying beneath. Alternative SEM and SIM in combination with site-specific in situ FIB sample sectioning made it possible to image the submicrometre structures of gland epithelium cells with reproducibility, repeatability and in the same range of magnifications as in transmission electron microscopy (TEM). At the present state of technology, ultrastructural elements imaged by the FIB /SEM system cannot be directly identified by comparison with TEM images.

Effect of Cathode Microstructure on the Cathode Polarization in the Sintered Strontium-Doped Lanthanum Manganite/Yttria Stabilized Zirconia Solid Oxide Fuel Cells

“…As a consequence, the negative charge built up on the surface of the specimen generates a negative electrical field [69,70]. This electrical field will interfere with the collection of secondary electrons and, even worse, deflect the incident beam and damage the specimen [70][71][72]. In low voltage SEM, although surface potential was reduced by decreasing the number of incident electrons on the surface, a high conductivity coating should be applied on the specimen.…”

“…High quality images are achieved by increasing secondary electron yield. The regular C and Au-Pd coatings will not meet these stringent requirements and may introduce image artifacts [70,71]. In the recently developed variable pressure SEM, charge at surface is neutralized by positive ions introduced by gas.…”

“…In the recently developed variable pressure SEM, charge at surface is neutralized by positive ions introduced by gas. This gas, however, reduces the efficiency of collecting secondary electrons [70,73].…”

Focused ion beam/scanning electron microscopy characterization of cell behavior on polymer micro-/nanopatterned substrates: A study of cell–substrate interactions

“…A combination of complementary information, gathered by classical secondary electron images (obtained from the interaction of the sample with a primary electron beam) and novel ion images (obtained from the interaction of the sample with a primary ion beam), can be obtained. Initial results of the combination of these techniques can be found in a recent study of the imaging of cross-sectional views of biological specimens (Drobne et al, 2005). However, to our knowledge, the combination of FIB and SEM techniques has not been applied to the study of cell-substrate However, in order to obtain faithful results using these techniques, high quality sample preparation procedures are essential.…”

Nano-Tomography of Porous Geological Materials Using Focused Ion Beam-Scanning Electron Microscopy

“…This is nowadays routinely performed in the semiconductor industry to investigate the performance of devices [24]. In addition to traditional biological and materials sciences [25][26][27][28][29][30][31][32][33], FIB-SEM analysis of natural solid materials, e.g., geological materials, has recently become increasingly popular [34][35][36][37].…”