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Journal of Biomedical Optics

Focused ion beam for microscopy and in situ sample preparation: application on a crustacean digestive system
Author(s): Damjana Drobne; Marziale Milani; Monica Ballerini; Alexis Zrimec; Maja Berden Zrimec; Francesco Tatti; Kazimir Draslar
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Paper Abstract

We prove that the focused ion beam (FIB) machine can be used as a microscope and as an in situ cutting device for tissue and cells. For the first time we obtain high-resolution ion images, complemented by electron imaging of different animal tissues both from uncoated and coated samples. In our study, we select the digestive system of Porcellio scaber (isopoda, crustacea) as a test system for FIB microscopy and in situ sample preparation. After the milling operation, some of the ultrastructural elements of hepatopancreatic cells can clearly be recognized. Also, FIB operation reveals significant differences in structural integrity between the apical and basal parts of hepatopancreatic cells, which have not been observed before by classical microscopy techniques. FIB microscopy and in situ sample preparation have advantages over classical microscopy techniques because of: 1. in situ site-specific 2-D sectioning and imaging of subsurface microstructures; 2. no need to embed the sample prior to sectioning; and 3. a wide range of magnifications while imaging the same sample.

Paper Details

Date Published: 1 November 2004
PDF: 6 pages
J. Biomed. Opt. 9(6) doi: 10.1117/1.1803846
Published in: Journal of Biomedical Optics Volume 9, Issue 6
Show Author Affiliations
Damjana Drobne, Univ. v Ljubljana (Slovenia)
Marziale Milani, Univ. degli Studi di Milano-Bicocca (Italy)
Monica Ballerini, Univ. degli Studi di Milano-Bicocca (Italy)
Alexis Zrimec, Institute of Physical Biology (Slovenia)
Maja Berden Zrimec, Institute of Physics Biology (Slovenia)
Francesco Tatti, FEI Italia (Italy)
Kazimir Draslar, Univ. of Ljubljana (Slovenia)

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