Tomography of insulating biological and geological materials using focused ion beam (FIB) sectioning and low-kV BSE imaging.
Tomography in a focused ion beam (FIB) scanning electron microscope (SEM) is a powerful method for the characterization of three-dimensional micro- and nanostructures. Although this technique can be routinely applied to conducting materials, FIB-SEM tomography of many insulators, including biological, geological and ceramic samples, is often more difficult because of charging effects that disturb the serial sectioning using the ion beam or the imaging using the electron beam. Here, we show that automatic tomography of biological and geological samples can be achieved by serial sectioning with a focused ion beam and block-face imaging using low-kV backscattered electrons. In addition, a new ion milling geometry is used that reduces the effects of intensity gradients that are inherent in conventional geometry used for FIB-SEM tomography.
De Winter
D A Matthijs
DA
Electron Microscopy and Structure Analysis, Cellular Architecture and Dynamics, Faculty of Sciences, Utrecht University, NL-3584 CH Utrecht, The Netherlands.
Schneijdenberg
C T W M
CT
Lebbink
M N
MN
Lich
B
B
Verkleij
A J
AJ
Drury
M R
MR
Humbel
B M
BM
D002478:Cells, Cultured; D042783:Endothelial Cells / Q000648:ultrastructure*; D004730:Endothelium, Vascular / Q000166:cytology*; D006801:Humans; D008274:Magnesium; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D008867:Microtomy; D017655:Silicon Compounds / Q000737:chemistry*; D016610:Tissue Embedding / Q000379:methods; D016707:Tissue Fixation / Q000379:methods; D014054:Tomography / Q000379:methods*; D014470:Umbilical Cord / Q000098:blood supply / Q000166:cytology
D017655:Silicon Compounds; C503823:forsterite; D008274:Magnesium