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Focused Ion Beam/Scanning Electron Microscopy Studies of Porcellio scaber (Isopoda, Crustacea) Digestive Gland Epithelium Cells

Damjana Drobne, Marziale Milani,* Alexis Zrimec,† Maja Berden Zrimec,† Francesco Tatti,‡ Kazimir Dras(breve)lar

Department of Biology, University of Ljubljana, Ljubljana, Slovenia; *Materials Science Department, University of Milano-Bicocca, Milano, Italy; †Institute of Physical Biology, Grosuplje, Slovenia; ‡FEI Italia, Rome, Italy

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Summary: The focused ion beam (FIB) was used to prepare cross sections of precisely selected regions of the digestive gland epithelium of a terrestrial isopod P. scaber (Isopoda, Crustacea) for scanning electron microscopy (SEM). The FIB / SEM system allows ad libitum selection of a region for gross morphologic to ultrastructural investigation, as the repetition of FIB/SEM operations is unrestricted. The milling parameters used in our work proved to be satisfactory to produce serial two-dimensional (2-D) cuts and/or three-dimensional (3-D) shapes on a submicrometer scale. A final, cleaning mill at lower ion currents was employed to minimize the milling artifacts. After cleaning, the milled surface was free of filament- and ridge-like milling artifacts. No other effects of the cleaning mill were observed.

Key words: focused ion beam, scanning electron microscopy, ultramicroscopy, sample preparation, digestive glands, terrestrial isopod Porcellio scaber

PACS: 07.78.+s, 61.16.Bq, 87.59.-e, 87.64.Dz

This research was funded by the Ministry of Education, Science and Sport of the Government of the Republic of Slovenia, through grants Nos. S39-431-001/12945/99 and J1-3186, and supported by the Foundation for Advances in Medicine and Science, Inc. (FAMS, Inc.).