Zuber Benoît, Nikonenko Irina, Klauser Paul, Muller Dominique, Dubochet Jacques
Laboratoire d'Analyse Ultrastructurale, Bâtiment de Biologie, Université de Lausanne, CH-1015 Lausanne, Switzerland.
Proc Natl Acad Sci U S A. 2005 Dec 27;102(52):19192-7. doi: 10.1073/pnas.0509527102. Epub 2005 Dec 14.
Cryo-electron microscopy of vitreous section makes it possible to observe cells and tissues at high resolution in a close-to-native state. The specimen remains hydrated; chemical fixation and staining are fully avoided. There is minimal molecular aggregation and the density observed in the image corresponds to the density in the object. Accordingly, organotypic hippocampal rat slices were vitrified under high pressure and controlled cryoprotection conditions, cryosectioned at a final thickness of approximately 70 nm and observed below -170 degrees C in a transmission electron microscope. The general aspect of the tissue compares with previous electron microscopy observations. The detailed analysis of the synapse reveals that the density of material in the synaptic cleft is high, even higher than in the cytoplasm, and that it is organized in 8.2-nm periodic transcleft complexes. Previously undescribed structures of presynaptic and postsynaptic elements are also described.
玻璃体切片的冷冻电子显微镜技术能够在接近自然状态下以高分辨率观察细胞和组织。标本保持水合状态,完全避免了化学固定和染色。分子聚集极少,图像中观察到的密度与物体中的密度相对应。因此,将大鼠海马器官型切片在高压和可控冷冻保护条件下进行玻璃化处理,切成最终厚度约为70纳米的冷冻切片,并在透射电子显微镜下于零下170摄氏度以下进行观察。该组织的总体外观与先前的电子显微镜观察结果相符。对突触的详细分析表明,突触间隙中的物质密度很高,甚至高于细胞质中的密度,并且这些物质以8.2纳米的周期性跨间隙复合物形式组织排列。还描述了以前未描述过的突触前和突触后元件的结构。