Moriguchi K, Higuchi N, Murakami Y, Yoshimura F, Nakamura H, Ohno N
Department of Anatomy, School of Dentistry, Aichi-Gakuin University, Nagoya, Aichi 464-8650, Japan.
Biotech Histochem. 2003 Apr;78(2):129-33. doi: 10.1080/10520290310001593784.
We used a rapid freezing and freeze-substitution technique without glutaraldehyde and OsO4 fixation for the electron microscopic immunocytochemical demonstration of the surface structure of Bacteroides forsythus, an anaerobic Gram-negative periodontopathogen. Cells were applied to pieces of filter paper and freeze-substituted by plunge-freezing in liquid propane, substituted in methanol containing 0.5% uranyl acetate, and infiltrated with LR White resin. The membrane ultrastructure of B. forsythus was preserved well, and the labeling density of the freeze-submitted cells was compared to a conventional processing method. Our results show the usefulness of the freeze-substitution method for immunohistochemical studies of B. forsythus.
我们采用了一种无需戊二醛和四氧化锇固定的快速冷冻和冷冻置换技术，用于电子显微镜免疫细胞化学显示福赛斯坦纳菌（一种厌氧革兰氏阴性牙周病原体）的表面结构。将细胞置于滤纸片上，通过投入液氮中速冻进行冷冻置换，在含有0.5%醋酸铀的甲醇中进行置换，并用LR White树脂渗透。福赛斯坦纳菌的膜超微结构保存良好，并将冷冻置换细胞的标记密度与传统处理方法进行了比较。我们的结果表明冷冻置换法对福赛斯坦纳菌的免疫组织化学研究有用。
