Scanning Probe Lithography in BioNanoTechnology

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Scanning Probe Lithography in BioNanoTechnology

• Department of Mechanical Engineering and
  Materials Science
• Woo-Kyung Lee

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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)
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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)
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Atomic Force Microscope (AFM)
1. System
2. Scanner
3. Cantilever
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Nanoshaving and Nanografting

• The simplest technique using self-assembly
  technique
• Thiolated SAM and gold surface

Liu et al., Acc.Chem.Res. 2000,33,457
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Examples
BSA
BSA
Lysozyme
Abell et al., Langmuir 2003, 19, 10557
Liu et al., PNAS 2002, 99, 5165
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Examples
Zauscher et al., Nanoletters 2004, 4, 373
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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., Science 283, 661 (1999)
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Examples
Protein Nanostructures (BSA)
Direct DPN of Oligonucleotides
Mirkin et al., Science 2002, 296, 1836
Zauscher et al., Nanoletters 2002, 2, 1203
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Anodization Lithography
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Enzymatic Lithography
Dan Luo, Materials Today, Nov. 2003
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Examples

• Staphylococcal serine V8 protease immobilization
  on AFM cantilever and Peptides surfaces
• Biotinylated peptide

Miyake et al., Nanoletters 2003, 3, 1471
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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 37C for 1 hour.

Chilkoti et al., JACS, accepted
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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