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TEM Observations of Bio-Conjugated Streptavidin-Gold Nanoparticles

Published online by Cambridge University Press:  01 February 2011

Ai Leen Koh  and
Robert Sinclair
Ai Leen Koh
Affiliation:
alkoh@stanford.edu, Stanford University, Materials Science and Engineering Department, Bldg 550, 416 Escondido Mall, Stanford, CA, 94305, United States, 650-353-0086
Robert Sinclair
Affiliation:
bobsinc@stanford.edu, Stanford University, Materials Science and Engineering Department, Stanford, CA, 94305, United States
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Abstract

Simultaneous visualization of inorganic nanoparticles and their bio-functionalized coating under the TEM can be achieved with negative staining, where electron dense material is introduced onto the specimen to enhance contrast. Using this technique, streptavidin-functionalized gold nanoparticles were negatively stained using phosphotungstic acid (PTA) at pH 7.0 and then observed using the TEM. The proteins appear as light regions/halos surrounding the gold nanoparticles, with widths similar to those of streptavidin protein molecules reported in literature. Experiments were performed to bio-conjugate streptavidin-gold nanoparticles to biotinylated antibodies and biotinylated actin, and then negatively staining the samples with PTA for TEM analyses. In the first experiment, an increase in halo widths in more than 60% of the particles was observed. In the second experiment, the nanoparticles were found to localize around the long actin filaments. Negative staining is useful for studying bio-functionalized nanoparticles as it enables the proteins surrounding the nanoparticles to be imaged and studied using the TEM.

Keywords

transmission electron microscopy (TEM)biologicalnanostructure
Type
Research Article
Information
MRS Online Proceedings Library (OPL) , Volume 1019: Symposium FF – Engineered Nanoscale Materials for the Diagnosis and Treatment of Disease , 2007 , 1019-FF05-01
Copyright
Copyright © Materials Research Society 2007

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