Buckley I K, Porter K R
J Microsc. 1975 Jul;104(2):107-20. doi: 10.1111/j.1365-2818.1975.tb04010.x.
To determine the overall fine structure of whole, unsectioned cells, cells from rat embryos were cultured on Formvar, glutaraldehyde/osmium-fixed, transferred to grids, dehypdrated, critical point dried, then examined by transmission electron microscopy at either 80 or 1000 kV. In contrast to air-dried material, critical point dried cells revealed each component clearly and with excellent contrast. All normal cytoplasmic structures (including coated vesicles, polyribosomes, microtubules and other fine components) were readily identifiable. Extensive structures such as microtubules and the endoplasmic reticulum (which appear fragmented in sections) were well displayed. At 1000 kV the beam readily penetrated even the thick nuclear and perinuclear cell regions and produced exceptionally crisp images. The methods described provide a simplified approach to the study of overall cell fine structure.
为了确定完整、未切片细胞的整体精细结构，将大鼠胚胎细胞培养在福尔马林中，经戊二醛/锇固定，转移到网格上，脱水，临界点干燥，然后在80千伏或1000千伏下用透射电子显微镜检查。与空气干燥的材料相比，临界点干燥的细胞能清晰地显示每个成分，对比度极佳。所有正常的细胞质结构（包括被膜小泡、多核糖体、微管和其他精细成分）都很容易识别。微管和内质网等广泛结构（在切片中看起来是破碎的）得到了很好的展示。在1000千伏下，电子束甚至能轻易穿透细胞核和核周细胞的厚区域，并产生异常清晰的图像。所描述的方法为研究细胞整体精细结构提供了一种简化方法。
