Pawley J B, Walther P, Shih S J, Malecki M
Integrated Microscopy Resource, Madison, WI 53706.
J Microsc. 1991 Feb;161(Pt 2):327-35. doi: 10.1111/j.1365-2818.1991.tb03093.x.
Recent advances in the design of the scanning electron microscope (SEM) column, such as the coupling of a field-emission gun to a low-aberration immersion lens and the availability of a high-stability cryo-transfer stage, make low-temperature, low-voltage SEM (LTLVSEM) possible at very high resolution. We have used this combination to obtain results with uncoated biological specimens. The trichocyst from a Paramecium was used as a test specimen to observe the shrinkage of this structure as the temperature is raised from 170 K to room temperature following freeze-drying. High-magnification stereo images were obtained of trichocysts that had been prepared by freezing, freeze-substitution and critical-point drying and which were subsequently viewed by LTLVSEM to reduce beam damage and contamination.
扫描电子显微镜（SEM）柱设计方面的最新进展，例如场发射枪与低像差浸没透镜的耦合以及高稳定性低温传输台的可用性，使得在非常高的分辨率下进行低温、低电压SEM（LTLVSEM）成为可能。我们利用这种组合对未镀膜的生物标本进行了研究。来自草履虫的刺丝泡被用作测试标本，以观察在冷冻干燥后温度从170 K升至室温时该结构的收缩情况。通过冷冻、冷冻置换和临界点干燥制备的刺丝泡获得了高放大倍率的立体图像，随后通过LTLVSEM观察以减少电子束损伤和污染。
