Specimen Preparation of the Human Cerebellar Cortex for Scanning Electron Microscopy Using a t-Butyl Alcohol Freeze-Drying Device

Authors

Teruyuki Hojo, University of Occupational and Environmental Health, JapanFollow

Abstract

A freeze-drying device was applied to t-butyl alcohol substituted nerve cells, fibers and synaptic terminals. Ten percent formalin-fixed human cerebellar cortex specimens were postfixed in 2.5% glutaraldehyde in 0.1 M cacodylate buffer solution and were rinsed three times in 5% sucrose solution in the same buffer. After the postfixed specimens were dehydrated using a graded series of ethanols and then transferred into a graded series of t-butyl alcohols (freezing point 25.4°C), the t-butyl alcohol substituted specimens were freeze-dried at 15°C and at high vacuum (5x10^-2 Torr). The freeze-dried specimens were sputter coated with gold. Scanning electron microscopy revealed synaptic terminals on the surfaces of a Golgi cell and a small flat polygonal cell. Rough somatic surfaces of granule cells were also observed.

Recommended Citation

Hojo, Teruyuki (1996) "Specimen Preparation of the Human Cerebellar Cortex for Scanning Electron Microscopy Using a t-Butyl Alcohol Freeze-Drying Device," Scanning Microscopy: Vol. 1996: No. 10, Article 27.
Available at: https://digitalcommons.usu.edu/microscopy/vol1996/iss10/27