A New Drying Method of Plant Specimens for Scanning Electron Microscopy the t-Butyl Alcohol Freeze-Drying Method

The t-butyl alcohol freeze-drying method (lnoue and Osatake 1988) is a simple drying method of biological materials for scanning electron microscopy Fixed specimens were immersed in t-butyl alcohol after dehydration throgh a graded series of ethanol. Specimens in the alcohol were then forzen in a refrigerator. They were placed in the bell jar of a vacuum evaporator and simply evacuated with a rotary pump. The samples were completely dried within 40–60 min after the frozen alcohol was sublimated in the vacuum, when the specimen was examined by scanning electron microscopy (SEM), both surface and intracellular structures were demonstrated in three-dimension without any significant drying artifacts. Careful comparison of the results indicated that the SEM imayes obtained by this method were either superior or equal to those obtained by the critical point drying method.