Wolosewick J J, Porter K R
J Cell Biol. 1979 Jul;82(1):114-39. doi: 10.1083/jcb.82.1.114.
The cytoplasmic ground substance of cultured cells prepared for high voltage transmission electron microscopy (glutaraldehyde/osmium fixed, alcohol or acetone dehydrated, critical-point dried) consists of slender (3-6 nm Diam) strands--the microtrabeculae (55)--that form an irregular three-dimensional lattice (the microtrabecular lattice). The microtrabeculae interconnect the membranous and nonmembranous organelles and are confluent with the cortices of the cytoplast. The lattice is found in all portions of the cytoplast of all cultured cells examined. The possibility that the lattice structure is an artifact of specimen preparation has been tested by (a) subjecting whole cultured cells (WI-38, NRK, chick embryo fibroblasts) to various chemical (aldehydes, osmium tetroxide) and nonchemical (freezing) fixation schedules, (b) examination of model systems (erythrocytes, protein solutions), (c) substantiating the relaibility of critical-point drying, and (d) comparing images of whole cells with conventionally prepared (plastic-embedded) cells. The lattice structure is preserved by chemical and nonchemical fixation, though alterations in ultrastructure can occur especially after prolonged exposure to osmium tetroxide. The critical-point method for drying specimens appears to be reliable as is the freeze-drying method. The discrepancies between images of plastic-embedded and sectioned cells, and images of whole, critical-point dried cells appear to be related, in part, to the electron-scattering properties of the embedding resin. The described observations indicate that the microtrabecular lattice seen in electron micrographs closely represents the nonrandom structure of the cytoplasmic ground substance of living cultured cells.
为进行高压透射电子显微镜检查而制备的培养细胞(经戊二醛/锇固定、酒精或丙酮脱水、临界点干燥)的细胞质基质由细长的(直径3 - 6纳米)细丝——微梁(55)组成,这些细丝形成不规则的三维晶格(微梁晶格)。微梁将膜性和非膜性细胞器相互连接,并与细胞质体的皮质相融合。在所检查的所有培养细胞的细胞质体的所有部分都发现了这种晶格。通过以下方式测试了晶格结构是标本制备假象的可能性:(a)使整个培养细胞(WI - 38、NRK、鸡胚成纤维细胞)经受各种化学(醛类、四氧化锇)和非化学(冷冻)固定程序;(b)检查模型系统(红细胞、蛋白质溶液);(c)证实临界点干燥的可靠性;(d)将全细胞图像与传统制备(塑料包埋)的细胞图像进行比较。尽管特别是在长时间暴露于四氧化锇后可能会发生超微结构的改变,但化学和非化学固定都能保留晶格结构。用于干燥标本的临界点方法似乎与冷冻干燥方法一样可靠。塑料包埋和切片细胞的图像与整个临界点干燥细胞的图像之间的差异,部分似乎与包埋树脂的电子散射特性有关。所描述的观察结果表明,在电子显微照片中看到的微梁晶格紧密代表了活培养细胞细胞质基质的非随机结构。