Biointerface Engineering Hub capabilities

Report
ANFF-Vic
Biointerface Engineering Hub
@Swinburne
Interfacing biology with technology
About the facility
• Wide range of capabilities in surface
modification
• Thin films, polymer grafting, biomolecule
immobilisation
• Bioactive coatings in polymers, ceramics and
metals
Overview
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Plasma Polymerisation
Langmuir Blodgett Surface Coatings And Analysis
Multi Vessel Dip Coating
Dip-Pen Nanolithography (DPN)
Quartz Crystal Microbalance (QCM)
Mask Aligner
UV/Ozone Cleaner
Biochemistry and Cell Biology
Plasma Polymerisation
• Specifications
– 6 custom made reactors for plasma
polymerisation (PE-CVD)
– <1-100 nm thin, pinhole-free polymer
coatings
– Substrate independent
– Up to 20cm diameter samples
– Vacuum process (1 x10-2 mbar)
– Can be set up to etch, treat and deposit
surfaces
How Does It Work
Gaseous monomer introduced at reduced pressure
Molecules fragmented by application of RF voltage – ‘glow discharge’
Fragments react with surface and each other to form highly cross linked coatings
Monomer properties can be retained…control power and flow rate
Etch, treat, deposit
Chamber
Pump
P~10-2 mbar
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HF voltage
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Electrode
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Monomer
Gas
Air
Applications
• Chemistries
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Alkanes
Alcohols
Acids
Amines
Thiols
PEO-like
Fluorocarbons
NiPAAm
Anhydrides
Hydrophobic
Hydrophilic
Alkane bromides (ATRP Initiator)
Essential Oils (antimicrobial)
• Substrates
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Metals
Polymers
Ceramics
Semiconductors
Fibres
Patterning
PDMS
Examples
Cell viability uncoated fibres vs. acrylic acid
coated fibres in 3D
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100
90
Percentage Live
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Uncoated
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Coated
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10
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96 Hours
Time
Chemically Patterned Surfaces
Bioarrays, biomaterials, sensors,
adhesion promotion,
Surface modification of electrospun fibres
for improved cell growth
Key References and Examples:
http://dx.doi.org/10.1002/elps.200800619
http://dx.doi.org/10.1021/la902930z
http://onlinelibrary.wiley.com/doi/10.1002/ppap.200900040/abstract
Langmuir Blodgett
Surface Coatings And Analysis
• Specifications*
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KSV-NIMA LB trough (medium)
Surface Area (cm2)
Trough top inner dimensions (LxWxH mm)
Maximum compression ratio
Barrier speed (mm/min)
Balance measuring range (mN/m)
273
364x75x4
10.8
0.1...270
0...150
– Film formation on gas/liquid and liquid/liquid interface
– Film deposition on solid substrates
– Microscopy trough
*http://www.ksvnima.com/langmuir-and-langmuir-blodgett-troughs
http://www.atascientific.com.au/langmuir-blodgett-films.html
How Does LB Work
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Trough filled with water (respective liquid)
Surfactant/Lipid is applied on surface of the liquid
Barriers are moved inwards to reduce surface area of Liquid
Surfactant rearranges on Surface
Forces can be measured with a Wilhelmy plate
Substrate can be dipped in solution and pulled out for monolayer formation
*http://www.ksvnima.com/langmuir-and-langmuir-blodgett-troughs
Applications
– Surface Tension measurements
– Dip coating of substrates for
monolayer formation
– Lipid bilayer formation
– Surfactant characterisation
– Drug-membrane studies
– Cell-membrane interactions
Uses:
Membrane Perturbation Activity of Cationic
Phenylene Ethynylene Oligomers and
Polymers: Selectivity against Model
Bacterial and Mammalian Membranes
http://dx.doi.org/10.1021/la102269y
*http://www.ksvnima.com/langmuir-and-langmuir-blodgett-troughs
KSV-NIMA Multivessel Dipcoater
• Multiple dipping of single substrate in different solutions
• Sample:
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Weight (g)
Height (mm)
Width (mm)
Thickness (mm)
0...500
25...300
5...950
0.15...15
Dipping speed: 1-1000 mm/min
Up to 10 samples dipped simultaneously
Create layer-by-layer (LbL) coatings from polyelectrolytes
Assemble colloidal arrays
*http://www.ksvnima.com/$2/ksv-nima-dcbrochure.pdf
Layer-by-layer (LbL) of Polyelectrolytes
Science 29 August 1997:
Vol. 277 no. 5330 pp. 1232-1237
DOI: 10.1126/science.277.5330.1232
Dip-Pen Nanolithography (DPN)
• Specifications
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Nanoink Nscriptor
Direct writing of molecules on surfaces
Bottom-up fabrication
< 1µm patterning from supplied design
Integrates with photolithography processing
Image drawn on gold using alkane thiol ink
How Does DPN Work
• Uses experimental set up of Atomic Force
Microscopy (AFM)
• Three main components of DPN
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AFM-tip as “pen”
Sample surface as “paper”
Molecular chemicals with affinity to surface
as “ink”
• Molecules (ink) transported from tip to surface via capillary
transport trough formed water meniscus
Applications
• Higher resolution/finer structures (sub- 100 nm) than
traditional printing methods
• Selective patterning of different molecules at
different sites on surface
• DPN allows multiple patterning at multiple
scales/shapes/sizes
• Applications:
– Biosensors
– Cell engineering
Quartz Crystal Microbalance (QCM)
• Specifications
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Q-Sense E4
Number of sensors 4
Volume above each sensor ~ 40 μl
Minimum sample volume ~ 300 μl
Working temperature 15 to 65°C,
(software controlled)
– Typical flow rates 50-200 μl/min
– Frequency and Dissipation analysis
http://www.q-sense.com/q-sense-e4
http://www.atascientific.com.au/surface-thin-film-interactions.html
How Does QCM Work
• Quartz crystal vibrates at distinct
frequency
• Adsorbed molecules result in change of
frequency
• Adsorbed mass correlates with frequency
and can be quantified
• Dissipation correlates with rigidity of
coating
Applications for QCM
• Kinetic studies of self assembly processes, thin film formation, protein and
drug interactions,
• Viscoelastic behaviours of thin films and adsorbed/immobilised layers
Adsorption kinetics of lipid vesicles are displayed as
Δf(t) and ΔD(t) for vesicles extruded through 30 nm
pores. Adsorption curves are presented for (a) Si3N4,
(b) oxidized Pt, (c) TiO2, and (d) SiO2
http://pubs.acs.org/doi/full/10.1021/la0263920
Mask Aligner
• Specifications
– OAI Model 200 Mask Aligner and UV Exposure
System
– Housed in Class 1000 cleanroom
– Air bearing / vacuum chuck levelling system, the
substrate is leveled quickly and gently, for
parallel photo mask alignment and uniform
contact across the wafer during contact
exposure.
– The system is capable of one micron resolution
and alignment precision.
*http://www.oainet.com/oai-200pp.html
UV/Ozone Cleaner
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UV/Ozone cleaning of surfaces
Removal of organic residues from surfaces
Oxydation/Hydrophilisation of surfaces
Cross linking of UV-curable resins
Clean AFM tips
Oxidize PDMS microfluidic devices
Prepare surfaces prior to gluing
http://www.atascientific.com.au/surface-and-afm-tip-cleaner.html
How Does The UV Cleaner Work
• The samples are placed underneath UV lamp
• UV light reacts with ambient oxygen to form Ozone
• Ozone is a very reactive compound that can react
with a large number of organic materials to form
volatile molecules like CO2 which are released into
the environment
• UV light can also act as cross linker of alkene
compounds
Biochemistry and Cell Biology
• PC2 laboratories for bacterial and cell culture
• Wide range of biochemical assays available
– ELISA
– Western blotting
– Protein assays
– Cell viability assays
• Confocal and Epi-fluorescence Microscopy
Biointerface Engineering Hub
• Contacts:
– Prof Sally McArthur
• [email protected]
• 61 3 92148452
– Dr Thomas Ameringer
• [email protected]
• 61 3 92144970

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