scanning probe lithography in bionanotechnology department of mechanical engineering and materials...
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Scanning Probe Scanning Probe Lithography in Lithography in
BioNanoTechnologyBioNanoTechnology
Department of Mechanical Engineering Department of Mechanical Engineering and Materials Scienceand Materials Science
Woo-Kyung LeeWoo-Kyung Lee
Patterning Biomolecules on Patterning Biomolecules on SurfacesSurfaces
Applications:Diagnostic ImmunoassaysDNA Micro-arraysCell-ResearchBio sensorsCell culturing
Drive miniaturization:•Diffusive mass transport increases•Chemical reactions accelerated•Smaller sample volumes required•Stimuli presented to cell locally
Immobilization:Simplest form: PhysisorptionMore advanced: Covalent bond to substrateor high affinity ligand pairs
Non-specific Binding:Minimize indiscriminate protein adhesion through surface repulsion chemistry (PEO)
Getting SmallerGetting Smaller
100m spots produced with non-contact micro-arrayer(Perkin Elmer, 2001)
40 m 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)
Atomic Force Microscope (AFM)Atomic Force Microscope (AFM)
1. System 2. Scanner
3. Cantilever
Nanoshaving and NanograftingNanoshaving and Nanografting
The simplest The simplest technique using self-technique using self-assemblyassembly technique technique
Thiolated SAM and Thiolated SAM and gold surfacegold surface
Liu et al., Acc.Chem.Res. 2000,33,457
ExamplesExamples
Liu et al., PNAS 2002, 99, 5165 Abell et al., Langmuir 2003, 19, 10557
BSA
Lysozyme
BSA
ExamplesExamples
Zauscher et al., Nanoletters 2004, 4, 373
Dip-Pen NanolithographyDip-Pen Nanolithography (DPN) (DPN)
Piner et al., Science 283, 661 (1999)
Direct write, positive printing technique using alkane thiols as diffusive inks. Feature size depends on: tip radius, substrate roughness, contact time, humidity (meniscus size).
ExamplesExamples
2 m
Mirkin et al., Science 2002, 296, 1836 Zauscher et al., Nanoletters 2002, 2, 1203
Direct DPN of Oligonucleotides Protein Nanostructures (BSA)
Anodization LithographyAnodization Lithography
vapor phase monomerIn a stream of N2
ODMSSAMOxide Layer
Si
(a)
Oxide LayerSi
SiO
O
O
(c)Ph
Ru
PCy3
PCy3
ClCl
Si
Oxide LayerSi
O
O
O
(d)
Ph
Si
Oxide LayerSi
O
O
OCHCH3
n
(e)
VOxide LayerSi
(b)
1 2 3 4 5
1μm
(a)
1 2 3 4 5
He
igh
t (n
m)
Enzymatic LithographyEnzymatic Lithography
Dan Luo, Materials Today, Nov. 2003
ExamplesExamples
Staphylococcal serine V8 protease immobilization on AFM cantilever and Peptides surfaces
Biotinylated peptide
Miyake et al., Nanoletters 2003, 3, 1471
ExamplesExamples
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
ChallengeChallenge
Enzyme immobilized AFM cantileverEnzyme immobilized AFM cantileverIn-situ digestion of DNA SAMIn-situ digestion of DNA SAMDNA modification using specific enzymesDNA modification using specific enzymes
ProblemsProblems Activity of immobilized enzymesActivity of immobilized enzymes Control reaction temperature and timeControl reaction temperature and time
SolutionSolution Search for suitable enzyme-DNA systemSearch for suitable enzyme-DNA system