Department of Biology, Biotechnical Faculty, University of Ljubljana, Vecna pot 111, SI-1000, Ljubljana, Slovenia.
Protoplasma. 2010 Oct;246(1-4):41-8. doi: 10.1007/s00709-010-0118-8. Epub 2010 Feb 19.
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.
聚焦离子束（FIB）和扫描电子显微镜（SEM）常用于材料科学领域，用于对材料进行成像和分析。在过去的十年中，组合式 FIB/SEM 系统已被证明也适用于生命科学。我们已经研究了聚焦离子束/扫描电子显微镜系统在研究模式生物 Porcellio scaber（甲壳纲：等足目）生物组织方面的潜力。组织取自消化腺，按常规 SEM 或透射电子显微镜（TEM）的方式进行制备。将样品转移到 FIB/SEM 中进行 FIB 铣削和成像操作。FIB 铣削区域进行二次电子成像、背散射电子成像或能量色散 X 射线（EDX）分析。我们的结果表明，FIB/SEM 能够同时研究样品的宏观形态、细胞表面特征和亚表面结构。用 EDX 对相同的 FIB 暴露区域进行分析，以提供基本的成分数据。当样品按 TEM 方式制备时，FIB/SEM 获得的信息与 TEM 获得的信息相当，尽管放大倍数有限。成像、微操作和成分分析的组合似乎特别适用于研究上皮组织，上皮组织会受到各种影响其结构和功能的内源性和外源性条件的影响。FIB/SEM 是一种有前途的工具，可用于在正常、应激或病理条件下对上皮组织进行全面检查。
