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Scanning Probe Lithography in BioNanoTechnology

Scanning Probe Lithography in BioNanoTechnology. Department of Mechanical Engineering and Materials Science Woo-Kyung Lee. Patterning Biomolecules on Surfaces. Applications: Diagnostic Immunoassays DNA Micro-arrays Cell-Research Bio sensors Cell culturing. Drive miniaturization:

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Scanning Probe Lithography in BioNanoTechnology

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  1. Scanning Probe Lithography in BioNanoTechnology Department of Mechanical Engineering and Materials Science Woo-Kyung Lee

  2. Patterning Biomolecules on Surfaces • Applications: • Diagnostic Immunoassays • DNA Micro-arrays • Cell-Research • Bio sensors • Cell culturing • Drive miniaturization: • Diffusive mass transport increases • Chemical reactions accelerated • Smaller sample volumes required • Stimuli presented to cell locally • Immobilization: • Simplest form: Physisorption • More advanced: Covalent bond to substrate • or high affinity ligand pairs • Non-specific Binding: • Minimize indiscriminate protein adhesion • through surface repulsion chemistry (PEO)

  3. Getting Smaller 100 mm spots produced with non-contact micro- arrayer (Perkin Elmer, 2001) 40 mm biotin spots fabricated on PFP- activated polymer-COOH surfaces via microcontact printing (Chilkoti, 2001) 300 nm spot size IgG nanoarray generated by DPN (Mirkin, 2002)

  4. Atomic Force Microscope (AFM) 1. System 2. Scanner 3. Cantilever

  5. Nanoshaving and Nanografting • The simplest technique using self-assembly technique • Thiolated SAM and gold surface Liu et al., Acc.Chem.Res. 2000,33,457

  6. Examples BSA BSA Lysozyme Abell et al., Langmuir 2003, 19, 10557 Liu et al.,PNAS 2002, 99, 5165

  7. Examples Zauscher et al.,Nanoletters 2004, 4, 373

  8. Dip-Pen Nanolithography (DPN) Direct write, positive printing technique using alkane thiols as diffusive inks. Feature size depends on: tip radius, substrate roughness, contact time, humidity (meniscus size). Piner et al., Science283, 661(1999)

  9. 2 mm Examples Protein Nanostructures (BSA) Direct DPN of Oligonucleotides Mirkin et al., Science 2002, 296, 1836 Zauscher et al., Nanoletters 2002, 2, 1203

  10. P C y 3 C l R u C l (a) (b) P C y 3 ODMS SAM V Oxide Layer Oxide Layer Si Si CHCH3 P P h h Height (nm) O (c) (d) O 1 O 2 3 4 5 O Oxide Layer Oxide Layer Si Si O Si O n (a) O (e) Si O O vapor phase monomer In a stream of N2 Oxide Layer Si Si 1 2 3 5 4 1μm Anodization Lithography

  11. Enzymatic Lithography Dan Luo, Materials Today, Nov. 2003

  12. Examples • Staphylococcal serine V8 protease immobilization on AFM cantilever and Peptides surfaces • Biotinylated peptide Miyake et al., Nanoletters 2003, 3, 1471

  13. Examples • DNASE-I and single strand DNA surface • DNASE-I is physically patterned by DPN technique. • After DPN the substrate is incubated in reaction buffer for enzyme at 37°C for 1 hour. Chilkoti et al.,JACS, accepted

  14. Challenge • Enzyme immobilized AFM cantilever • In-situ digestion of DNA SAM • DNA modification using specific enzymes • Problems • Activity of immobilized enzymes • Control reaction temperature and time • Solution • Search for suitable enzyme-DNA system

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