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
Abell et al., Langmuir 2003, 19, 10557

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

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

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