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Nanometer Scale Protein Templates for Bionanotechnology Applications
KTH, School of Engineering Sciences (SCI), Applied Physics, Nanostructure Physics.
2005 (English)Doctoral thesis, comprehensive summary (Other scientific)
Abstract [en]

Nanofabrication techniques were used to manufacture nanometer scale protein templates. The fabrication approach employs electron beam lithography (EBL) patterning on poly(ethylene glycol) (PEG) thiol (CH3O(CH2CH2O)17NHCO(CH2)2SH) self-assembled monolayers (SAM) on Au. The PEG SAM prevented protein surface adhesion and binding sites for protein were created in the SAM by EBL. Subsequent to EBL, the patterns in the PEG SAM were backfilled with 40-nm NeutrAvidin-coated fluorescent spheres (FluoSpheres). The spontaneous and directed immobilization of the spheres from a solution to the patterns resulted in high resolution protein patterns. The FluoSpheres could be arranged in any arbitrary pattern with ultimately only one or a few FluoSpheres at each binding site.

Growth dynamics and SAM morphology of PEG on Au were studied by atomic force microscopy (AFM). PEG SAMs on three types of Au with different microstructure were examined: thermally evaporated granular Au and two types of Au films produced by hydrogen flame annealing of granular Au, Au(111) and "terraced" Au (crystal orientation unknown). The different Au surfaces' substructure affected the morphology and mechanical properties of the PEG SAM. On Au(111), AFM imaging revealed monolayer formation through three distinct steps: island nucleation, island growth, and coalescence. The fine-structure of the SAM revealed dendritic island formation - an observation which can be explained by attractive intermolecular interactions and diffusion-limited aggregation. Island growth was not observed on the "terraced" Au.

AFM studies of EBL patterned PEG SAMs on Au(111) revealed two different patterning mechanisms. At low doses, the pattern formation occurs by SAM ablation in a self-developing process where the feature depth is directly dose dependent. At higher doses electron beam induced deposition of material, so-called contamination writing, is seen in the ablated areas of the SAM. The balance between these two mechanisms is shown to depend on the geometry of the pattern.

In addition to PEG SAMs, fibronectin monolayers on SiO2 surfaces were patterned by EBL. The areas exposed with EBL lose their functionality and do not bind anti-fibronectin. With this approach we constructed fibronectin templates and used them for cell studies demonstrating pattern dependent cell geometries and cell adhesion.

Place, publisher, year, edition, pages
Stockholm: KTH , 2005. , xv, 108 p.
Series
Trita-FYS, ISSN 0280-316X ; 2005:65
Keyword [en]
nanotechnology, bionanotechnology, nanobiotechnology, electron beam lithography, EBL, poly(ethylene glycol), PEG, self-assembled monolayer, SAM, self-immobilization, protein pattern
National Category
Physical Sciences
Identifiers
URN: urn:nbn:se:kth:diva-530ISBN: 91-7178-178-1 (print)OAI: oai:DiVA.org:kth-530DiVA: diva2:14312
Public defence
2005-12-09, FA32, AlbaNova, Roslagstullsbacken 21, Stockholm, 10:15
Opponent
Supervisors
Note
QC 20101008Available from: 2005-12-02 Created: 2005-12-02 Last updated: 2012-03-21Bibliographically approved
List of papers
1. Poly(ethylene glycol) Self-Assembled Monolayer Island Growth
Open this publication in new window or tab >>Poly(ethylene glycol) Self-Assembled Monolayer Island Growth
2005 (English)In: Langmuir, ISSN 0743-7463, E-ISSN 1520-5827, Vol. 21, 2981-2987 p.Article in journal (Refereed) Published
Abstract [en]

Here, we report a study of the morphology and growth dynamics of a self-assembled monolayer (SAM) of the amide containing poly(ethylene glycol) (PEG) thiol (CH3O(CH2CH2O)(17)NHCO(CH2)(2)SH) on atomically flat Au(111) surfaces. SAM growth from a 20 mu M ethanolic solution reveals island growth through three distinct steps: island nucleation, island growth, and coalescence. The coalescence-step, filling voids in the SAM, is by far slowest. The fine structure study reveals dendritic island formation, an observation which can be explained by attractive intermolecular interactions and surface diffusion-limited aggregation. We have also observed a change in the island height, which peaks during the island growth phase. This height change can be associated with a molecular conformational transition.

Keyword
sum-frequency spectroscopy, size distribution, surface modification, protein adsorption, hydrogen-bonds, in-situ, gold, alkanethiols, microscopy, mechanisms
National Category
Physical Sciences
Identifiers
urn:nbn:se:kth:diva-8845 (URN)10.1021/la0471792 (DOI)000228042400052 ()2-s2.0-16244376824 (Scopus ID)
Note
QC 20101008Available from: 2005-12-02 Created: 2005-12-02 Last updated: 2017-12-14Bibliographically approved
2. Substrate Microstructure Influences Properties of Poly(ethylene glycol) Monolayers on Au
Open this publication in new window or tab >>Substrate Microstructure Influences Properties of Poly(ethylene glycol) Monolayers on Au
(English)In: Nano Letters, ISSN 1530-6992Article in journal (Other academic) Submitted
National Category
Physical Sciences
Identifiers
urn:nbn:se:kth:diva-8846 (URN)
Note
QC 20101008Available from: 2005-12-02 Created: 2005-12-02 Last updated: 2010-10-08Bibliographically approved
3. Directed Immobilization of Protein-Coated Nanospheres toNanometer-Scale Patterns Fabricated by Electron Beam Lithographyof Poly(ethylene glycol) Self-Assembled Monolayers
Open this publication in new window or tab >>Directed Immobilization of Protein-Coated Nanospheres toNanometer-Scale Patterns Fabricated by Electron Beam Lithographyof Poly(ethylene glycol) Self-Assembled Monolayers
2006 (English)In: Langmuir, ISSN 0743-7463, E-ISSN 1520-5827, Vol. 22, no 11, 5100-5107 p.Article in journal (Refereed) Published
Abstract [en]

Controlling the spatial organization of biomolecules on solid supports with high resolution is important for a widerange of scientific and technological problems. Here we report a study of electron beam lithography (EBL) patterningof a self-assembled monolayer (SAM) of the amide-containing poly(ethylene glycol) (PEG) thiol CH3O(CH2CH2O)17-NHCO(CH2)2SH on Au and demonstrate the patterning of biomolecular features with dimensions approaching 40 nm.The electron dose dependence of feature size and pattern resolution is studied in detail by atomic force microscopy(AFM), which reveals two distinct patterning mechanisms. At low doses, the pattern formation occurs bySAMablationin a self-developing process where the feature size is directly dose-dependent. At higher doses, electron beam-induceddeposition of material, so-called contamination writing, is seen in the ablated areas of the SAM. The balance betweenthese two mechanisms is shown to depend on the geometry of the pattern. The patterned SAMs were backfilled withfluorescent 40-nm spheres coated with NeutrAvidin. These protein-coated spheres adhered to exposed areas in theSAM with high selectivity. This direct writing approach for patterning bioactive surfaces is a fast and efficient wayto produce patterns with a resolution approaching that of single proteins.

Keyword
scanning force microscopy, chemical nanolithography, deposited tips, surfaces, silicon, adsorption, biosensor, resists
National Category
Physical Sciences
Identifiers
urn:nbn:se:kth:diva-8847 (URN)10.1021/la052306v (DOI)000237593000035 ()2-s2.0-33745736728 (Scopus ID)
Note
Uppdaterad från "In press" till published: 20101008. Tidigare titel: Directed Protein Adhesion to Nanometer Scale Patterns Fabricated by Electron Beam Lithography on Poly(ethylene glycol) Self-Assembled Monolayers QC 20101008Available from: 2005-12-02 Created: 2005-12-02 Last updated: 2017-12-14Bibliographically approved

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