Dahl R, Staehelin L A
Research Service, VA Medical Center, Denver, Colorado 80220.
J Electron Microsc Tech. 1989 Nov;13(3):165-74. doi: 10.1002/jemt.1060130305.
The two main advantages of cryofixation over chemical fixation methods are the simultaneous stabilization of all cellular components and the much faster rate of fixation. The main drawback pertains to the limited depth (less than 20 microns surface layer) to which samples can be well frozen when freezing is carried out under atmospheric conditions. High-pressure freezing increases the depth close to 0.6 mm to which samples can be frozen without the formation of structurally distorting ice crystals. This review discusses the theory of high-pressure freezing, the design of the first commercial high-pressure freezing apparatus (the Balzers HPM 010), the operation of this instrument, the quality of freezing, and novel structural observations made on high-pressure-frozen cells and tissues.
与化学固定方法相比，冷冻固定的两个主要优点是能同时稳定所有细胞成分以及固定速度快得多。主要缺点是在大气条件下进行冷冻时，样品能被良好冷冻的深度有限（表层小于20微米）。高压冷冻可将样品能被冷冻而不形成结构扭曲冰晶的深度增加到接近0.6毫米。本文综述了高压冷冻的理论、第一台商用高压冷冻设备（Balzers HPM 010）的设计、该仪器的操作、冷冻质量以及对高压冷冻细胞和组织的新结构观察。
