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六甲基二硅氮烷(HMDS)、Peldri II以及临界点干燥法用于生物样本扫描电子显微镜观察的比较

Comparison of hexamethyldisilazane (HMDS), Peldri II, and critical-point drying methods for scanning electron microscopy of biological specimens.

作者信息

Bray D F, Bagu J, Koegler P

机构信息

Department of Biological Sciences, University of Lethbridge, Alberta, Canada.

出版信息

Microsc Res Tech. 1993 Dec 15;26(6):489-95. doi: 10.1002/jemt.1070260603.

Abstract

Three different drying methods, critical-point drying (CPD), Peldri II, and hexamethyldisilazane (HMDS), were compared using representative animal (rat kidney, trachea, duodenum, lung, and red blood cells) and plant (leaves from ten species of monocotyledons and dicotyledons) specimens. All three drying methods produced identical results with animal specimens. Plant specimens showed signs of shrinkage regardless of which drying method was employed. The order of preservation quality from best to worst for leaves was CPD > Peldri II > HMDS, with the CPD method providing substantially better results in all but one case. Postfixation of leaves with osmium tetroxide resulted in poorer preservation in all instances. Peldri II caused complete extraction of leaf cuticular wax, while both both CPD and HMDS showed minimal extraction compared with samples air dried directly from acetone. These results indicate that HMDS provides a time-saving and inexpensive alternative to CPD for animal specimens. Plant specimens, particularly those containing cells with large central vacuoles, are adequately preserved only with the CPD method. In addition, postfixation with osmium should be avoided when processing plant specimens for scanning electron microscopy.

摘要

使用代表性的动物(大鼠肾脏、气管、十二指肠、肺和红细胞)和植物(十种单子叶植物和双子叶植物的叶子)标本,比较了三种不同的干燥方法,即临界点干燥(CPD)、Peldri II和六甲基二硅氮烷(HMDS)。对于动物标本,所有三种干燥方法都产生了相同的结果。无论采用哪种干燥方法,植物标本都出现了收缩迹象。叶片保存质量从最好到最差的顺序是CPD > Peldri II > HMDS,除了一个案例外,CPD方法在所有情况下都提供了明显更好的结果。用四氧化锇对叶片进行后固定在所有情况下都会导致保存效果较差。Peldri II导致叶片表皮蜡完全被提取,而与直接从丙酮中空气干燥的样品相比,CPD和HMDS都显示出最少的提取。这些结果表明,对于动物标本,HMDS为CPD提供了一种省时且廉价的替代方法。植物标本,特别是那些含有具有大中央液泡细胞的标本,只有用CPD方法才能得到充分保存。此外,在处理用于扫描电子显微镜的植物标本时,应避免用锇进行后固定。

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