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Examination of Large Unilamellar Vesicles (Luvs) Using Cryo-hrsem and Cryo-Stem

Published online by Cambridge University Press:  02 July 2020

K. L. Caran ,
R. P. Apkarian  and
F. M. Menger
K. L. Caran
Affiliation:
Department of Chemistry, Emory University, 1515 Pierce Drive, Atlanta, GA30322 Integrated Microscopy & Microanalytical Facility, Department of Chemistry, Emory University, 1515 Pierce Drive, Atlanta, GA30322
R. P. Apkarian
Affiliation:
Integrated Microscopy & Microanalytical Facility, Department of Chemistry, Emory University, 1515 Pierce Drive, Atlanta, GA30322
F. M. Menger
Affiliation:
Department of Chemistry, Emory University, 1515 Pierce Drive, Atlanta, GA30322
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Extract

Vesicles derive their structure from the aqueous environment surrounding and within them. It follows that traditional EM specimen preparation techniques of fixation, dehydration and drying can induce a variety of morphological changes to the membranes. Vitrification of aqueous suspensions of vesicles provides a means for observation of these structures in their fully hydrated unfixed state. Common cryogenic techniques for the visualization of these and other colloidal particles include cryo-TEM of vitrified thin films and freeze-fracture TEM (FF-TEM) of platinum replicas. We report the use of cryo-high resolution SEM (cryo- HRSEM) and cryo-STEM for the study of the morphology of membrane features of synthetic extruded vesicles.

LUVs were prepared by the extrusion method. Dried lipid films of l-palmitoyl-2-oleoyl-phosphatidylcholine (POPC) were hydrated with 0.5 mL of Milli-Q water and stirred for 10 minutes to make suspensions (10 mM in lipids) which were extruded 19 times through a polycarbonate filter with 100 nm pores.

Type
Biopolymers and Biomemetics
Information
Microscopy and Microanalysis , Volume 5 , Issue S2: Proceedings: Microscopy & Microanalysis '99, Microscopy Society of America 57th Annual Meeting, Microbeam Analysis Society 33rd Annual Meeting, Portland, Oregon August 1-5, 1999 , August 1999 , pp. 1210 - 1211
Copyright
Copyright © Microscopy Society of America

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References

1.Almgreen, M.et al., Current Opinion in Colloid and Interface Science 1(1996)270CrossRefGoogle Scholar
2.Tahara, Y. and Fujiyoshi, Y., Micron 25:2(1994)141CrossRefGoogle Scholar
3.Gustaffson, J. et al., Biochimica et Biophysica Acta 1235(1995)305CrossRefGoogle Scholar
4.Dorovska-Taran, V. et al., Analytical Biochemistry 240(1996)37CrossRefGoogle Scholar
5.Bergmeier, M. et al., J Colloid and Interface Science 203(1998)1CrossRefGoogle Scholar
6.Gradzielski, M. et al., J Phys Chem B 101:10(1997)1719CrossRefGoogle Scholar
7.Apkarian, R.P. et al., Microscopy and Microanalysis (1998, submitted)Google Scholar
8.Apkarian, R.P. et al., Proc Ann Meeting MSA 54(1996)914Google Scholar
9. This research was supported by an NIH grant to F.M. Menger.Google Scholar