CIL:1446

Rabbit psoas skeletal muscle fibers were held at constant length and rigor induced by immersion in glycerol. The fibers were quick-frozen by contact with a liquid helium- cooled copper block in a Heuser-type cryopress, and freeze-etched in a Balzers 400 freeze fracture machine. The sample was then rotary-replicated with Pt-C at 24 degrees and visualized in a JEOL 100CX2 transmission electron microscope operated at 100 kev. Images were recorded on Kodak 4489 film at 68,000x magnification. Stereo pairs were taken at +5 and -5 degrees. Films were scanned with a 20 micrometer pixel spacing on a Nikon Coolscan 9000ED scanner. Stereo pair with CIL:1447将兔腰大肌骨骼肌纤维维持于恒定长度，并通过浸入甘油诱导肌肉僵直。随后将纤维置于Heuser型冷冻压榨仪 (Heuser-type cryopress) 中，通过与液氦冷却的铜块接触实现快速冷冻。样本经Balzers 400型冷冻断裂仪 (Balzers 400 freeze fracture machine) 完成冷冻蚀刻后，在24°条件下以铂-碳 (Pt-C) 进行旋转复型。随后使用工作电压为100 keV的JEOL 100CX2型透射电子显微镜 (JEOL 100CX2 transmission electron microscope) 进行观测，图像以68000倍放大倍率记录于柯达4489胶片上。拍摄了±5°倾角的立体对图像，采用尼康Coolscan 9000ED扫描仪以20微米的像素间距对胶片进行扫描，该立体对对应编号CIL:1447。