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Morphological Characterization of an Elastin-Mimetic Triblock Copolymer Utilizing CRYO-HRSEM

Published online by Cambridge University Press:  02 July 2020

E.R. Wright ,
V.P. Conticello  and
R.P. Apkarian
E.R. Wright
Affiliation:
Department of Chemistry, Emory University,Atlanta, GA, 30322
V.P. Conticello
Affiliation:
Integrated Microscopy and Microanalytical Facility, Department of Chemistry, Emory University, Atlanta, GA, 30322
R.P. Apkarian
Affiliation:
Department of Chemistry, Emory University,Atlanta, GA, 30322
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Abstract

The elastin-mimetic tri-block copolymer, C5, with the polypeptide repeat [((VPAVG)(IPAVG)3(VPAVG))m((VPGVG)2(VPGEG)(VPGVG)3)n((VPAVG)(IPAVG)3(VPAVG))m] was produced via protein hyper-expression utilizing a pET plasmid and Escherichia coli (BL21- Gold(DE3)) as the bacterial host. The gene coding for the polypeptide was produced by a multimerization protocol that has undergone recent modifications. A sample of the elastinmimetic block copolymer, C5, was prepared as a 20 to 25 percent protein solutions in sterile ddH20. Approximately 10 μL of the solution was pipetted into 3 mm gold planchets (Balzers BU 012 130T), which had been pre-equilibrated to 4°C in an isothermal environmental cooler (Torrey Pines Scientific). The temperature of the cooler was raised to 20°C and allowed to stabilize for 10 minutes. The solidified samples, were plunge-frozen in liquid ethane at its melting point (−183°C); the samples were stored in liquid nitrogen (LN2). A sample was transferred to and mounted on the precooled (∽−170°C) Oxford CT-3500 cryo-stage held in the cryo-preparation chamber.

Type
Advances in Imaging Techniques for Biomaterlals (Organized by S. Eppel)
Information
Microscopy and Microanalysis , Volume 7 , Issue S2: Proceedings: Microscopy & Microanalysis 2001, Microscopy Society of America 59th Annual Meeting, Microbeam Analysis Society 35th Annual Meeting, Long Beach, California August 5-9, 2001 , August 2001 , pp. 136 - 137
Copyright
Copyright © Microscopy Society of America 2001

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References

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