Scanning Probe Lithography in BioNanoTechnology - PowerPoint PPT Presentation

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Scanning Probe Lithography in BioNanoTechnology

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... 40 mm biotin spots fabricated on PFP- activated polymer-COOH surfaces via microcontact printing (Chilkoti, 2001) 300 nm spot size IgG nanoarray generated by ... – PowerPoint PPT presentation

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Title: Scanning Probe Lithography in BioNanoTechnology


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., Science 283, 661 (1999)
9
Examples
Protein Nanostructures (BSA)
Direct DPN of Oligonucleotides
Mirkin et al., Science 2002, 296, 1836
Zauscher et al., Nanoletters 2002, 2, 1203
10
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 37C 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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