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Protoplasma

, Volume 246, Issue 1–4, pp 41–48 | Cite as

Focused ion beam (FIB)/scanning electron microscopy (SEM) in tissue structural research

  • Vladka Lešer
  • Marziale Milani
  • Francesco Tatti
  • Živa Pipan Tkalec
  • Jasna Štrus
  • Damjana Drobne
Original Article

Abstract

The focused ion beam (FIB) and scanning electron microscope (SEM) are commonly used in material sciences for imaging and analysis of materials. Over the last decade, the combined FIB/SEM system has proven to be also applicable in the life sciences. We have examined the potential of the focused ion beam/scanning electron microscope system for the investigation of biological tissues of the model organism Porcellio scaber (Crustacea: Isopoda). Tissue from digestive glands was prepared as for conventional SEM or as for transmission electron microscopy (TEM). The samples were transferred into FIB/SEM for FIB milling and an imaging operation. FIB-milled regions were secondary electron imaged, back-scattered electron imaged, or energy dispersive X-ray (EDX) analyzed. Our results demonstrated that FIB/SEM enables simultaneous investigation of sample gross morphology, cell surface characteristics, and subsurface structures. The same FIB-exposed regions were analyzed by EDX to provide basic compositional data. When samples were prepared as for TEM, the information obtained with FIB/SEM is comparable, though at limited magnification, to that obtained from TEM. A combination of imaging, micro-manipulation, and compositional analysis appears of particular interest in the investigation of epithelial tissues, which are subjected to various endogenous and exogenous conditions affecting their structure and function. The FIB/SEM is a promising tool for an overall examination of epithelial tissue under normal, stressed, or pathological conditions.

Keywords

Histopathology Hepatopancreas Terrestrial isopods Crustacea EDX analyses 

Abbreviations

FIB

Focused ion beam

SEM

Scanning electron microscopy

TEM

Transmission electron microscopy

EDX

Energy dispersive X-ray

BSE

Back-scattered electrons

Notes

Acknowledgments

We thank FEI Italy for the access to Strata DB235 M at the University of Modena e Reggio Emilia, Italy. This work was supported by the Slovenian Ministry of Higher Education, Science and Technology (grants J1-6473 and 3311-03-831253), World Federation of Scientists, and the Slovenian Science Foundation (WFS National Scholarship SZF-V.Lešer/WFS-01/2007). We thank Bill Millne for the English editing.

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Vladka Lešer
    • 1
  • Marziale Milani
    • 2
  • Francesco Tatti
    • 3
  • Živa Pipan Tkalec
    • 1
  • Jasna Štrus
    • 1
  • Damjana Drobne
    • 1
  1. 1.Department of Biology, Biotechnical FacultyUniversity of LjubljanaLjubljanaSlovenia
  2. 2.Materials Science Department and Laboratory FIB/SEM “Bombay”University of Milano-BicoccaMilanItaly
  3. 3.FEI ItaliaMilanItaly

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