Specimen preparation of the human cerebellar cortex for scanning electron microscopy using a t-butyl alcohol freeze-drying device.
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 degrees C), the t-butyl alcohol substituted specimens were freeze-dried at 15 degrees C and at high vacuum (5 x 10(-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.
Hojo
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Department of Anatomy and Anthropology, School of Medicine, University of Occupational and Environmental Health, Kitakyushu City, Japan. hojo@med.uoeh-u.ac.jp
D000368:Aged; D000369:Aged, 80 and over; D000440:Butanols; D002525:Cerebellar Cortex / Q000648:ultrastructure*; D005612:Freeze Drying; D016591:Histocytological Preparation Techniques; D006801:Humans; D008297:Male; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D017729:Presynaptic Terminals / Q000648:ultrastructure; D016707:Tissue Fixation
D000440:Butanols