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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

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