Thomas Giddings (2011) CIL:14676, Oscillatoria tenuis. CIL. Dataset

The thylakoids of the vegetative cells of the filamentous cyanobacteria, Oscillatoria tenuis, are capable of oxygen-evolving photosynthesis. Freeze-fracture replicas of the vegetative cells revealed the structure of the thylakoids.

The process of freeze-fracture splits memebranes along their hydrophobic interior revealing two complementary fracture faces designated P-face and E-face. Each fracture faces has a population of particles which represent intramembrane proteins or protein complexes.

Oscillatoria cultures were maintained on agar plates. Cells were collected and suspended in 10 mL dH2O, spun at 2,000 g for 10 min in a clinical centrifuge, resuspended in 5 mL dH2O. Added 5 mL 70% glycerol over 1 hr with continuous stirring, spun at 2,000 g for 10 min in a clinical centrifuge. Pellets were placed on copper support discs, frozen in liquid Freon 12 and stored under liquid nitrogen. Replicas were prepared on a Blazers BA 360 freeze-etch unit and examined in a JEOL TEM 100C at 50,000X magnification.

蓝藻丝状体 Oscillatoria tenuis 的营养细胞叶绿体能够进行氧演化光合作用。冷冻断裂复制品揭示了营养细胞叶绿体的结构。冷冻断裂技术沿细胞膜疏水内部进行分裂，揭示了两个互补的断裂面，分别称为 P 面和 E 面。每个断裂面都存在一组代表跨膜蛋白或蛋白复合物的颗粒。Oscillatoria 的培养在琼脂平板上进行。收集细胞并悬浮于 10 mL 重蒸水中，在临床离心机中以 2,000 g 的速度离心 10 分钟，然后重新悬浮于 5 mL 重蒸水中。在 1 小时内加入 5 mL 70% 的甘油，并持续搅拌，然后在临床离心机中以 2,000 g 的速度离心 10 分钟。将沉淀物放置在铜支持盘上，在液态 Freon 12 中冷冻，并在液氮下储存。复制品在 Blazers BA 360 冷冻蚀刻装置上制备，并在 JEOL TEM 100C 透射电子显微镜下以 50,000 倍的放大倍数进行观察。