Uwins P J, Murray M, Gould R J
Centre for Microscopy and Microanalysis, University of Queensland, St. Lucia, Australia.
Microsc Res Tech. 1993 Aug;25(5-6):412-8. doi: 10.1002/jemt.1070250510.
Four common scanning electron microscope (SEM) processing techniques involving freeze-substitution and chemical fixation were compared with fresh unprocessed samples imaged in an environmental scanning electron microscope (ESEM) using small pieces of potato tubers as test specimens. Potato tubers were chosen for this investigation because of their high moisture content and, consequently, the common need for extensive processing for conventional, high vacuum SEM imaging. ESEM results showed that the fresh unprocessed specimens were essentially unaltered, showing clear potato cell structure, morphology, and cell content. However, processed samples showed strong differences to the fresh samples: freeze-substituted specimens showed fine networks of material stretching across the surface of cells. These structures may represent fibrillar material or may be artifact caused during processing. Chemical fixation almost entirely destroyed the microstructure of these potato samples.
以小块马铃薯块茎为测试样本,将四种涉及冷冻置换和化学固定的常见扫描电子显微镜(SEM)处理技术与在环境扫描电子显微镜(ESEM)中成像的新鲜未处理样本进行了比较。选择马铃薯块茎进行这项研究是因为它们的含水量高,因此通常需要进行大量处理才能用于传统的高真空SEM成像。ESEM结果表明,新鲜未处理的样本基本未发生变化,呈现出清晰的马铃薯细胞结构、形态和细胞内含物。然而,处理后的样本与新鲜样本表现出很大差异:冷冻置换后的样本显示出横跨细胞表面延伸的精细物质网络。这些结构可能代表纤维状物质,也可能是处理过程中产生的假象。化学固定几乎完全破坏了这些马铃薯样本的微观结构。