Robinow C F, Marak J
J Cell Biol. 1966 Apr;29(1):129-51. doi: 10.1083/jcb.29.1.129.
The structure and mode of division of the nucleus of budding yeast cells have been studied by phase-contrast microscopy during life and by ordinary microscopy after Helly fixation. The components of the nucleus were differentially stained by the Feulgen procedure, with Giemsa solution after hydrolysis, and with iron alum haematoxylin. New information was obtained in cells fixed in Helly's by directly staining them with 0.005% acid fuchsin in 1% acetic acid in water. Electron micrographs have been made of sections of cells that were first fixed with 3% glutaraldehyde, then divested of their walls with snail juice, and postfixed with osmium tetroxide. Light and electron microscopy have given concordant information about the organization of the yeast nucleus. A peripheral segment of the nucleus is occupied by relatively dense matter (the "peripheral cluster" of Mundkur) which is Feulgen negative. The greater part of the nucleus is filled with fine-grained Feulgen-positive matter of low density in which chromosomes could not be identified. Chromosomes become visible in this region under the light microscope at meiosis. In the chromatin lies a short fiber with strong affinity for acid fuchsin. The nucleus divides by elongation and constriction, and during this process the fiber becomes long and thin. Electron microscopy has resolved it into a bundle of dark-edged 150 to 180 A filaments which extends between "centriolar plaques" that are attached to the nuclear envelope.
通过相差显微镜在活体状态下以及在经赫利固定后用普通显微镜对出芽酵母细胞核的结构和分裂方式进行了研究。细胞核的组成成分通过福尔根染色法、水解后用吉姆萨溶液染色以及用铁明矾苏木精进行了差异染色。在用1%醋酸水溶液中的0.005%酸性品红直接对赫利固定的细胞进行染色后,获得了新的信息。已对先用3%戊二醛固定、然后用蜗牛酶去除细胞壁并再用四氧化锇后固定的细胞切片制作了电子显微镜照片。光学显微镜和电子显微镜给出了关于酵母细胞核组织结构的一致信息。细胞核的外周部分被相对致密的物质(蒙德库尔的“外周簇”)占据,该物质对福尔根染色呈阴性。细胞核的大部分充满了低密度的细颗粒福尔根阳性物质,在其中无法识别出染色体。在减数分裂时,在光学显微镜下该区域的染色体可见。在染色质中有一条对酸性品红有强烈亲和力的短纤维。细胞核通过伸长和缢缩进行分裂,在此过程中该纤维变得细长。电子显微镜已将其解析为一束边缘深色的150至180埃的细丝,这些细丝在附着于核膜的“中心粒斑”之间延伸。