An application of scanned focused ion beam milling to studies on the internal morphology of small arthropods

Abstract

Summary For the first time a scanned focused ion beam of approximately 50 nm diameter has been used to prepare biological material. Small defined areas of the surface were removed by ion etching to allow examination of the underlying structures with a scanning electron microscope. Different milling procedures were carried out on two anatomical features in mites of the genus Halarachne (Halarachnidae: Mesostigmata). In the first, square holes were milled into the surface of the peritrematal plate to reveal the structure of the underlying respiratory peritrematal groove. In the second, transverse cuts were made across the shafts of the sensory sensilli which make up the sensory Haller's organ on tarsus I. This latter procedure revealed detail both within the core and walls of sensilli. Details of specimen preparation and milling procedures, as well as suitability and interpretation of results, are presented.

Focused ion beam/scanning electron microscopy studies of Porcellio scaber (Isopoda, Crustacea) digestive gland epithelium cells

“…After the final cleaning mill, the milled surface was smooth. Similar milling irregularities appearing in the form of striations were also observed by Young et al (1993) when milling the cuticular structures of arthropods. Low-energy ions are also used to remove the presence of thin layers on the exposed surface as a result of melting, implantation of Ga + ions, or redeposition of milled material (Haswell et al 2003).…”

Insight into <i>Emiliania huxleyi</i> coccospheres by focused ion beam sectioning

“…FIB-SEM sectioning was shown to be appropriate for biological samples for the first time in 1993 (Young et al, 1993). Since this seminal study this method has improved and several groups have reported about the use of FIB-SEM microscopes for material sciences and biological materials (Phaneuf, 1999;Uchic et al, 2006;Giannuzzi et al, 2007;Leser et al, 2009;Grandfield and Engquvist, 2012;Srot et al, 2012).…”

The Correlation between Ion Beam/Material Interactions and Practical FIB Specimen Preparation

“…In addition to TEM specimen preparation, FIB milling techniques have also been effective in producing specimens for subsequent analysis by scanning electron microscopy~SEM!, secondary ion mass spectrometry~SIMS!, scanning transmission electron microscopy~STEM!, AUGER electron spectroscopy~AES!, and atomic force microscopy~AFM; Mackenzie et al, 1993Mackenzie et al, , 1994Young et al, 1993a;Huffman et al, 1994;Ishitani et al, 1995;Kamino et al, 1998;Prenitzer et al, 1998;Stevie et al, 1999Stevie et al, , 2001Vartuli et al, 1999!. It should be noted that the fundamental concepts of ion-solid interactions presented in this work are equally applicable to any such potential application.…”