Freeze substitution of high-pressure frozen samples: the visibility of biological membranes is improved when the substitution medium contains water
Freeze substitution of high-pressure frozen samples: the visibility of biological membranes is improved when the substitution medium contains water
复制标题
DOI：
10.1046/j.1365-2818.2002.01064.x
复制
发表时间：
2002-10-01
影响因子：
2
通讯作者：
Ziegler, A
中科院分区：
文献类型：
--
作者：
Walther, P;Ziegler, A
生物膜经过高压冷冻和冷冻取代后，在电子显微镜下往往很难看到。样品和替代介质的含水量是影响膜可视性的重要因素之一。为了研究这种影响，我们用高压冷冻酵母细胞、大鼠胰腺组织和节肢动物组织分别在替代培养基中加水和不加水进行冷冻替代。如果替代介质中含有1-5%的水，生物膜的可见度一般会提高。这种效果在酵母细胞中尤其明显，在用无水培养基冷冻替代后，膜的可视性很差，而在替代培养基中加入5%的水后，膜的可视性很好。
Biological membranes are often poorly visible with the electron microscope after high-pressure freezing and freeze-substitution. The water content of the sample and of the substitution medium is one factor among others that strongly influences membrane visibility. In order to investigate this effect, high-pressure frozen yeast cells, rat-pancreas tissue and arthropod tissue were freeze-substituted with and without adding water to the substitution medium. The visibility of the biological membranes was generally improved if the substitution medium contained 1-5% water. The effect was especially pronounced in yeast cells, where membrane visibility was poor after freeze-substitution with water-free medium but good after addition of 5% water to the substitution medium.