一种新型冷冻样品台的构建及其在植物样品扫描电镜观测中的应用  

Construction of new frozen sample table and its application in scanning electron microscope observation of plant samples

作　　者：涂牧东 黄俊峰 周保范 张军伟[1] 邓霞[1] 李华[1,2] TU Mudong;HUANG Junfeng;ZHOU Baofan;ZHANG Junwei;DENG Xia;LI Hua(Electron Microscopy Centre,School of Materials and Energy,Lanzhou University,Lanzhou 730000,China;School of Chemistry and Chemical Engineering,Lanzhou University,Lanzhou 730000,China;School of Physical Science and Technology,Lanzhou University,Lanzhou 730000,China)

机构地区：[1]兰州大学材料与能源学院电镜中心,甘肃兰州730000 [2]兰州大学化学化工学院,甘肃兰州730000 [3]兰州大学物理科学与技术学院,甘肃兰州730000

出　　处：《实验技术与管理》2022年第8期28-32,37,共6页Experimental Technology and Management

基　　金：发光与实时分析化学教育部重点实验室(西南大学)开放基金(LAMS20210701);甘肃省科技计划项目(21JR7RA491,21JR7RA494)。

摘　　要：扫描电子显微镜(SEM)目前广泛应用于各种类型样品的观测,但在含有水分的植物样品的观测上还存在许多问题,比如植物中的水分会蒸发到电镜中,损坏电子枪;而且植物样品在高真空环境下直接观测,或采用干燥法来去除样品中的水分,都会使样品或多或少发生变形,从而无法得到原始形貌。该文研制了适用于各种常规SEM的新型冷冻样品台,并对其在植物细胞的SEM观测性能进行了研究探讨。对比采用生物样品常规制样方法(临界点干燥法)获得的拟南芥侧根细胞SEM图像,结果发现使用该样品台观测所获得的图像更加准确,避免了临界点干燥法制样引起的样品个别部位的收缩和细胞变形;并且使用该样品台,在高真空、高加速电压下对欧洲千里光幼茎皮层细胞进行观测,样品也表现出了足够的稳定性,没有因高电压发生损害,可满足植物样品较高分辨率的观察需求;使用该样品台,无需对现有常规SEM进行改装,制样简单快速,能有效地提高常规SEM对含有水分的有机生物样的测试能力。Scanning electron microscope(SEM) is widely used in the observation of various types of samples at present, but there are still many problems in the observation of plant samples containing water, such as the water in plants will evaporate into the electric mirror and damage the electron gun. Moreover, the direct observation of plant samples in high vacuum environment or the use of drying method to remove the water in the samples will make the samples more or less deformed, so the original morphology can not be obtained. In this paper, a new type of frozen sample table suitable for various conventional SEM is developed, and its SEM observation of plant cells is studied and discussed. Compared with the SEM images of Arabidopsis lateral root cells obtained by the conventional biological sample preparation method(critical point drying method), the results show that the images obtained by using this sample platform are more accurate and avoided the shrinkage and cell deformation of individual parts of the sample caused by the critical point drying method. The sample platform can also be used to observe the leaf cells of European Senecio under high vacuum and high accelerating voltage, it shows sufficient stability and is not damaged by high voltage, which can meet the observation needs of plant samples with high resolution. Using this sample table, there is no need to refit the existing conventional SEM, the sample preparation is simple and fast,which can effectively improve the testing ability of the conventional SEM for organic biological samples containing water.

关 键 词：扫描电子显微镜 样品台 植物样品 蓄冷材料 

分 类 号：Q336[生物学—遗传学]