CRITICAL-POINT DRYING VERSUS FREEZE-DRYING FOR SCANNING ELECTRON-MICROSCOPY - A QUANTITATIVE AND QUALITATIVE STUDY ON ISOLATED HEPATOCYTES
CRITICAL-POINT DRYING VERSUS FREEZE-DRYING FOR SCANNING ELECTRON-MICROSCOPY - A QUANTITATIVE AND QUALITATIVE STUDY ON ISOLATED HEPATOCYTES
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DOI：
10.1111/j.1365-2818.1985.tb02577.x
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发表时间：
1985-01-01
影响因子：
2
通讯作者：
ROSTGAARD, J
中科院分区：
文献类型：
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作者：
NORDESTGAARD, BG;ROSTGAARD, J
将临界点干燥和冷冻干燥作为扫描电子显微镜的制备方法进行了定性和定量的比较。以分离的[大鼠]肝细胞作为模型细胞。用Nomaski差示干涉显微镜对未固定、戊二醛固定、戊二醛+OsO_4固定、临界点干燥和冷冻干燥状态下的肝细胞进行了光镜观察。临界点干燥肝细胞收缩到戊二醛+OsO4固定体积的38%，而冷冻干燥肝细胞(冷冻于饱和氯仿水中，183℃冷冻干燥)。K=84h)收缩到戊二醛+OsO4固定体积的51%。临界点干燥细胞的透射和扫描电子显微镜显示细胞的超微结构和表面结构保存完好。冻干细胞的显微照片显示，内部冰晶破坏了超微结构，外部冰晶破坏了表面结构。双固定分离的肝细胞在缓冲液中储存时肿胀，临界点干燥后在储存过程中收缩。对于低放大倍率的扫描电子显微镜(最高可达.apprx3000倍)临界点干燥和冷冻干燥均可使用。对于高倍率扫描电子显微镜，临界点干燥优于冷冻干燥。
Critical-point drying and freeze drying were compared both quantitatively and qualitatively as preparative procedures for scanning electron microscopy [SEM]. Isolated [rat] hepatocytes were used as model cells. Nomaski differential interference contrast microscopy was used for light microscopic measurements of the hepatocytes in the unfixed, the glutaraldehyde fixed, the glutaraldehyde + OsO4 fixed, the critical-point dried and the freeze dried states. Critical point dried hepatocytes shrank to 38% of glutaraldehyde + OsO4 fixed volume, whereas optimal freeze dried hepatocytes (frozen in water saturated with chloroform and freeze dried at 183.degree. K for 84 h) shrank to 51% of glutaraldehyde + OsO4 fixed volume. Transmission and scanning electron micrographs of the critical-point dried cells showed well-preserved ultrastructure and surface structure. Micrographs of the freeze dried cells showed ultrastructure destroyed by internal ice crystals and surface structure destroyed by external ice crystals. Double-fixed isolated hepatocytes swelled during storage in buffer and shrank during storage after critical-point drying. For low magnification SEM (up to .apprx. 3000 times) both critical-point drying and freeze drying can be used. For high magnification SEM, critical-point drying is superior to freeze drying.