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利用环境扫描电子显微镜(ESEM)、低温扫描电子显微镜(Cryo-SEM)和激光扫描显微镜对挪威云杉早期体细胞胚胎进行成像。

Imaging of Norway spruce early somatic embryos with the ESEM, Cryo-SEM and laser scanning microscope.

作者信息

Neděla Vilém, Hřib Jiří, Havel Ladislav, Hudec Jiří, Runštuk Jiří

机构信息

Environmental Electron Microscopy Group, Institute of Scientific Instruments of the CAS, Brno, Czech Republic.

Environmental Electron Microscopy Group, Institute of Scientific Instruments of the CAS, Brno, Czech Republic.

出版信息

Micron. 2016 May;84:67-71. doi: 10.1016/j.micron.2016.02.011. Epub 2016 Mar 2.

Abstract

This article describes the surface structure of Norway spruce early somatic embryos (ESEs) as a typical culture with asynchronous development. The microstructure of extracellular matrix covering ESEs were observed using the environmental scanning electron microscope as a primary tool and using the scanning electron microscope with cryo attachment and laser electron microscope as a complementary tool allowing our results to be proven independently. The fresh samples were observed in conditions of the air environment of the environmental scanning electron microscope (ESEM) with the pressure from 550Pa to 690Pa and the low temperature of the sample from -18°C to -22°C. The samples were studied using two different types of detector to allow studying either the thin surface structure or material composition. The scanning electron microscope with cryo attachment was used for imaging frozen extracellular matrix microstructure with higher resolution. The combination of both electron microscopy methods was suitable for observation of "native" plant samples, allowing correct evaluation of our results, free of error and artifacts.

摘要

本文描述了挪威云杉早期体细胞胚(ESE)的表面结构,其作为一种具有异步发育的典型培养物。以环境扫描电子显微镜作为主要工具,并使用带低温附件的扫描电子显微镜和激光电子显微镜作为辅助工具,观察覆盖ESE的细胞外基质的微观结构,从而使我们的结果能够得到独立验证。在环境扫描电子显微镜(ESEM)的空气环境条件下,于550Pa至690Pa的压力以及样品-18°C至-22°C的低温下观察新鲜样品。使用两种不同类型的探测器对样品进行研究,以便研究薄表面结构或材料成分。带低温附件的扫描电子显微镜用于对冷冻的细胞外基质微观结构进行更高分辨率的成像。两种电子显微镜方法的结合适用于观察“天然”植物样品,从而能够对我们的结果进行正确评估,且无误差和假象。

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