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Field Emission Scanning Electron Microscopy Of Mouse Cerebellar Synaptic Contacts

Published online by Cambridge University Press:  02 July 2020

O.J. Castejón
Affiliation:
Instituto de Investigaciones Biológicas “Dres. Orlando J. Castejon y. Haydee Viloria de Castejón.” Facultad de Medicrna. Universidad del Zulia Maracaibo, Venezuela
R. P. Apkarian
Affiliation:
SEM Facilities. Chemistry Department. Emory University, Atlanta, GAUSA
H. V. Castejón
Affiliation:
Instituto de Investigaciones Biológicas “Dres. Orlando J. Castejon y. Haydee Viloria de Castejón.” Facultad de Medicrna. Universidad del Zulia Maracaibo, Venezuela
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Extract

Samples of albino mice cerebellar cortex were processed by the cryofracture method for scanning electron microscopy and examined with the field emission scanning electron microscope (FESEM). Albino mouse cerebellar cortex was excised, cut into 1-2 mm slices and inmersed in 4% glutaraldehyde in O. l M phosphate buffer solution, pH 7.4, for 24h at 4°C; and postfixed for 1 h in a similarly buffered 1% osmium tetroxide solution. Specimens were dehydrated in a graded serie of ethanol (30, 50, 70, 80, 90 2x100%) prior to wrapping individual tissue pieces in preformed absolute ethanol filled parafilm cryofracture packets. Rapid freezing of packets was performed by plunging into LN2. First, the packet was transferred from the LN2 storage vessel with LNT chilled forceps in order to avoid themial damage. Secondly, the cooled fracture blade was removed from the LN2, the packet was orientated under the blade, and immediately struck with a heavy tool.

Type
Biomedical Applications
Copyright
Copyright © Microscopy Society of America

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References

References:

1.Humphreys, W.J., Spurlock, B.O and Jonston, J.S., Stain Tech. 50 (1974) 119125.CrossRefGoogle Scholar
2.Apkarian, R.P., Caram, K.L. and Robinson, K.A., Microsc. Microanal. 5 (1999) 197207.CrossRefGoogle Scholar
3.Castejón, O.J. and Apkarian, R.P.. J. Subrracrosc. Cytol. Pathol. 24 (1992) 549562.Google Scholar
4. This research was supported by CONDES-LUZ. Maracaibo-Venezuela, and Chemistry Department. Emory University. Atlanta. Georgia, USAGoogle Scholar