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Harry HeinzelmannVP Nanotechnology & Life SciencesNanotechnology Tools for Life SciencesNeuchâtel, June 2009
Citation preview
Harry Heinzelmann
VP Nanotechnology & Life Sciences
Nanotechnology Tools for Life Sciences
Neuchâtel, June 2009
v1.19
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 1
CSEM profile
Privately held Innovation Center, incorporated, not for profit
since 1984, from watchmaking
about 70 shareholders (mostly private)
Privately held Innovation Center, incorporated, not for profit
since 1984, from watchmaking
about 70 shareholders (mostly private)
2008:
>65 Mio. CHF annual turnover, 395 employees
30 start-ups created since 2000
2008:
>65 Mio. CHF annual turnover, 395 employees
30 start-ups created since 2000
Activities:
Applied research (contract with Swiss Government)
Industrialization of technologies, product development
Activities:
Applied research (contract with Swiss Government)
Industrialization of technologies, product development
Technologies:
Micro- and Nanotechnology, Information Technology,
and System Engineering
Technologies:
Micro- and Nanotechnology, Information Technology,
and System Engineering
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 2
Bridge from Science to Innovation
CSEM profile
• research partners:
Applied
Res & Dev
Science &
Education
Basic Research
PhD programs
Teaching
Product
Development
Market
Success
Marketing
Sales
Customers
Industrialisation
• technologies
for innovations
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 3
Bridge from Science to Innovation
• wide range of technologies, large experience and network � innovative solutions
• highly qualified and experienced staff � fast developments
• IP portfolio to support the customers’ application � protected business
CSEM profile
• technologies for Green Solutions
• shareholders include
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 4
Technologies I (divisions in Neuchâtel)
CSEM profile
Microelectronics
Circuit Design, RF, Information Processing
Nanotechnology & Life Sciences
Optical and Bio MNT, Self-assembly, Sensors
Systems Engineering
Mechatronics, Signal Processing, Communication
Microsystems
MEMS, Cleanroom Infrastucture, Microscopy & Analysis
Time and Frequency
Atomic Clocks, Optical Advanced Systems
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 5
Technologies II (divisions outside Neuchâtel)
CSEM profile
Photonics (Zurich)
Image Sensing, Optoelectronics
Robotics (Alpnach)
Lab Automation, Packaging, Assembly
Thin Film Optics (Basel)
Optoelectronics, Replicated Optics
Nano Medicine (Landquart)
Imaging, Medical Sensors
CSEM UAE
CSEM Brazil
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 6
• extended experience in self-assembly of
polymer and nanoparticle systems
• block copolymer microphase separation
and copolymer lithography / MEMS
• molecular grafting chemistries, from and to
• controlled self-assembly of beads
• partnerships & projects:
• industrial collaborations:
Nanostructuring
Technologies and Applications
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 7
Top-Down vs. Bottom-Up
Nanostructuring
classical (micro-) fabrication
MEMS: Micro Electro Mechanical Systems
lithography:
VIS
UV, X-ray, e – beam
FIB (serial)
10
1 mm
1 µm
10
100
100
10
1 nm
1 Å
molecular self-assembly
“molecular nanotechnology” 4 µµµµm250 nm
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 8
Polymeric Self-Assembly I : Polymer Demixing
80 µm
50% PMMA / 50% PS 90% PMMA / 10% PS
5 µm
• demixing of immiscible polymer blends
• qualitative structures on the micron scale
• control over feature size and properties
• large variety of polymers available
Nanostructuring
• simple deposition technique
• selective solvent can remove one polymer type
• scalable to
large surfaces
� inexpensive and flexible method to control surface properties on a micron scale
large 5µm med 2µm small <1µm
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 9
Nanoporous Layers for Ink-jet Printer Paper
Nanostructuring – Polymer Demixing
Polymer paper
stable images
slow ink uptake, big spot size
Polymer layer
Cellulose
Nanoporous layer
(Alumina film)
Cellulose
Nanoporous paper
fast up-take, small spot size
image fading (light, gas,…)
paper, 5 µm x 5 µm
polymer film
transferred
on paper
polymer on Si
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 10
*patentpending
• nanoscale structures are
difficult to counterfeit,
and are mass-producible
Security Features for Anti-Counterfeiting Applications
Nanostructuring – Polymer Demixing
• market size for counterfeit goods (2004): 500 Bill. US$
for art pieces: >10 Bill. US$ (Europe)
• self-assembly structures are random and unique
• security features can be mass produced at low cost, both for mass id and unique fingerprints
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 11
�
Topography Gradients for Surface Interaction Screening
Blondiaux et al., submitted
PMMA / P2VP demixing on a pre-prepared surface chemistry gradient
Nanostructuring – Polymer Demixing
• surface coatings with controlled properties,
varying over short length scales
• combinatorial studies of cell-substrate interactions: effect of surface
roughness on cell adhesion and proliferation, with gradients adapted
to typical distances travelled by cells � study of cell locomotion
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 12
�
Polymeric Self-Assembly II : Microphase Separation
Nanostructuring
Krishnamoorthy et al., materials today (September 2006)
• Microphase Separation
• inexpensive & flexible method to generate
ordered structures on the molecular scale
• wide choice of functions and chemistries:
mechanical, chemical / catalytic,
optical, electrical, magnetic, …
10 -100 nm
-A-A-A- -B-B-B-
high
A-fraction
high
B-fraction
• block copolymer A-b-B
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 13
(Random) Nanostructures with Order and Function
Function:
• PI-b-PFS poly(isoprene-b-ferrocenylsilane)
• spincoating of 30nm thin film, plasma etch
• high density magnetic pattern: 4 1011 /cm2
Order:
• random and short range
• can be improved by templating
• topographical, chemical, temp, fields, …
Nanostructuring – Copolymer Microphase Separation
Fe
Si
CH3
CH3
H
n-Bu nm
PI-b-PFS
different FexOy stochiometries
from Korczagin, Vancso et al., Mesa+
PS-PFS
from Stoykovich et al., Science (2005)
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 14
�
Copolymer Lithography for Nano-Pillars and Nano-Pores
Nanostructuring – Copolymer Microphase Separation
Krishnamoorthy et al., Nanotechnology (2008)
etch mask from
copolymer patterns
from polymer
constituents with
different etch rates
in some cases it is
necessary to provide
an “amplification” of
the etch contrast
inverted micelles etch mask
RIE
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 15
�
Non-Wetting Surfaces with Nanopillar Structures
Nanostructuring – Copolymer Microphase Separation
Krishnamoorthy et al., Nanotechnology (2008)
planar SiNx silanised
with perfluorosilane:
contact angle 111°
Nanopillars in SiNx, 90nm high, 100nm periodicity
silanised with perfluorosilane:
water contact angle 150˚, highly mobile drop
• self-cleaning surfaces by functionalization
with perfluorosilane (wet or PVD)
transition from Wenzel to Cassie-Baxter wetting mode
for structure aspect ratio > 2:1
WCA adv 160° (compare to 110° on a flat surface)
hysteresis 5°, rolling angle 6°, 10ml droplet
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 16
Osmotic Biosensor based on Nanoporous Membranes
• nanoporous membranes from
copolymer lithography
• macro prototyping of osmotic sensor
• size selectivity supported specific
binding chemistry
Nanostructuring – Nanoporous Membranes
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 17
Wafer Scale Replication of Copolymer Lithography Patterns
Nanostructuring – Nanoporous Membranes
• replication by polymer casting
wafer scale PDMS casting
influence on:
• cell growth
• protein expression
• cytoskeleton organ.
• nanostructured surfaces for cell studies
• PMMA nanoporous membranes
• replication by embossing into PC foil
master by Ni electroplating
small medium large
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 18
BioMEMS for Nanotoxicity Tests
• experience in cell handling
dedicated infrastructure
• established knowledge in microfabrication
and replication technologies, in house fab
• nanotechnology / nanoparticle handling
• microfluidics design and prototyping
• partnerships & projects: InLiveTox
• industrial collaborations:
Technologies and Applications
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 19
Nanotoxicology – Risks of Nanoparticle Technology
• molecular nanotechnology “hype”
• “grey goo” & “green goo”
• new class of nano-materials with “unknown”
properties: carbon (CNT, buckyballs, …),
TiO2, SiO2, metallic (Au…), quantum dots
(CdS, CdSe, CdTe, etc.), polymeric…
BioMEMS
gold
latex
CNTs Catalytic CO Oxidation by a Gold
Nanoparticle, N. Lopez and J.K.
Norskov, J.Am.Chem.Soc.(2002)
• … in widespread applications: catalysts,
sunscreens, fuel cells, solar panels, …
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 20
Translocation Measurement Device – EU IP Nanosafe2
• problem: unknown effects of nanoparticles on human organisms
• microfabricated chip for the in vitro study of model epithelia transport properties
BioMEMS - Nanotoxicology
detection of nanoparticles
that cross the cell layer
porous Si3N4
membrane
confluent layer of
epithelial cells
electrodes for TEER
measurements
nanoparticle suspension
coming in
detection of inorganic nanoparticles off-line using inductively
coupled plasma mass spectrometry (ICP-MS)
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 21
On-Chip Electrical Characterization of Cell Layers
• microfabricated chip with cell culture wells
• porous membranes at the basis of each well
to allow toxins or drugs to pass through
• TransEpithelial Electrical Resistance (TEER)
to determine the tightness of a cell layer
BioMEMS - Nanotoxicology
• electrical contacts
• plastic holder
• glass support, to seal the
fluidic network
• PDMS fluidics
• SiN membrane
• PDMS in plastic holder,
electrical contacts at the bottom
Calu-3 cells grown in one of five wells
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 22
Intestinal, Liver & Endothelial NP Toxicity – InLiveTox
BioMEMS - Nanotoxicology
• CSEM, 4 university partners, Helmholtz Zentrum Berlin, Kirkstall Ltd, Alma
• objectives:
• develop in vitro test system to reduce/replace animal tests of nanoparticle toxicity
• replace the “lab rat”
by a setup of
• microfluidics and
• cell cultures
of model organs
• 3Rs: Replace, Reduce and Refine animal tests
• REACH: Registration, Evaluation, Authorisation & Restriction of Chemical Substances
‘Bloodstream’
Nanoparticles Sampling ports
‘Gastro Intestinal tract’ ‘Intestinal epithelium’
(co-culture of epithelial cells,
monocytes and dendritic cells)
‘Vascular endothelium’
(endothelial cells)
‘Liver’ (hepatocytes)
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 23
Probe Array Technology – PROBART
• speed up single probe operation
by parallel
imaging and
sensing
Nanotools
nisenet.org
• PROBART for Life Science applications, for
- bioarrays
- cells
but: operation in liquids!
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 24
Force Spectroscopy on Cells
• information about adhesion proteins,
cell mechanics, kinetics, …
• cell-surface, cell-cantilever, cell-cell
• meaningful only with sufficient
statistics, which makes experiments
rather tedious
• at current rate of a few cells per day,
not useful for screening formats
• array format would improve statistics
and make high throughput screening
formats more accessible
Nanotools – Probe Arrays
source: JPK
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 25
PROBART for Parallel Imaging
Nanotools – Probe Arrays
R lever
R ref
VEE (- 6V)Rlever
Rref
R1 R2
Vout
(~ 20 kohm)
4x4 array imaging in
buffer solution with
continuous zoom-in
probe
#6
probe
#13
probe
#15
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 26
Cell Adhesion Forces
Nanotools – Probe Arrays
Human osteoblasts, growing on
hemispherical pits (a, diameter 27 µm) and
nanopillars (b, 45nm high, replicated in a non-
metallic bone implant material
similar adhesion forces for cells in all
phases of the cell cycle (thus no need
for synchronization in future studies)
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 27
Nanotools for Ultimate Pipetting
• vast experience in Scanning Probe Methods
• MEMS design and fabrication in house
• fluidics design and fabrication
• surface chemistry and characterization
• experience in handling biomaterials,
nanoparticles in solutions
• partnerships & projects:
• industrial collaborations: first contacts with instrument makers
Technologies and Applications
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 28
Nanoscale Dispensing – NADIS
Nanotools – Nanoscale Dispensing
Nanoparticle suspensions
Materials for processing
Molecules in solution
• functional biomolecules for microarrays, such as
proteins or DNA
• metallic nanoparticles to form connects, catalyst
particles, optical and chemical functions, …
• etch resist materials, sol-gel precursors, …
deposition of liquidsin ultrasmall volumesfrom microscopic tips
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 29
�
NADIS with FIB Modified Probes
Nanotools – Nanoscale Dispensing
Meister et al., App.Phys.Lett. (2004)
1 µm
sub-attoliter volumes
• apertures with Ø down to 200 nm
• flexibility in location (off-center, …)
• possible to keep sharp AFM tips
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 30
Nanotools – Nanoscale Dispensing
0
0.5
1
0 2 4 6 8
Inte
nsit
y [a
.u.]
3 µm
applied pressure ~ 2mbar
NADIS of Fluorophores in Liquid Environments
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 31
NADIS for Liquid Exchange with Living Cells
Nanotools – Nanoscale Dispensing
• injection after perforation
of the cell membrane
• extraction of cytoplasm for
remote analysis
• towards patch clamping
viable neuroblastoma cells
Cell TrackerTM green staining
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 32
Nanostructuring
Conclusions
• polymer demixing
for random but regular microstructures
• co-polymer microphase separation
for well-arranged functional nanostructures and lithography
THANK YOU !
• collaborators from CSEM: AM Popa, M Klein, W Li, F Montage, R Pugin, …
• cleanroom team from COMLAB and CMI EPF Lausanne
• partners from U Mulhouse, U Twente, EPFL, …
Copyright 2009 I Nanotechnology & Life Sciences I Harry Heinzelmann I page 33
Nanotools
Conclusions
• probe array platform
for parallel force spectroscopy in biological environments
• nanoscale dispensing (NADIS)
for liquid arraying and cell manipulation
THANK YOU !
• collaborators from CSEM: J Przybylska, M Favre, J Polesel, A Meister, M Liley, …
• cleanroom team from COMLAB and CMI EPF Lausanne
• partners from IMT U Neuchâtel, U Lund, U Trento, ETHZ, EPFL, …
Thank you for your attention.