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Funded by the European Union - GA 737089 1Kick-Off Meeting, Barcelona, Jan 17-18, 2017
ChipScope project kick-off meeting
Lab-on-a-chip
Rudolf Heer, [email protected], AIT
Funded by the European Union - GA 737089 2Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Austrias biggest non-university research institute in numbers
Owners: 50.46% Republic of Austria
BMVIT Federal Ministry for Transport, Innovation and Technology
49.54% Federation of Austrian Industries
Employees: 1100 plus >250 on contract basis, thereof >100 PhD students
Financial Goal: 30% Cooperative Research
30% Contract Research
40% Basic Funding
Total Operating Income: >120 Mio. Euro
Funded by the European Union - GA 737089 3Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Funded by the European Union - GA 737089 4Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Diagnostic
Biosensors
Diagnostic
Biosensors
Business Unit Molecular Diagnostics
Saliva Diagnostics
Biomarker
Development
Assay
Development
Systems Integration
Point-of-Care
Bioinformatics
Funded by the European Union - GA 737089 5Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Diagnostic biosensor development
Funded by the European Union - GA 737089 6Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Biosensing- targets & physical transduction principles
Peptides(e.g. hormones)
Proteins(e.g. antibodies,
enzymes, hormones)
Nucleic acids(DNA, RNA,
mRNA, miRNA)
Viruses, cells
Biological targets
Waveguides
Surface plasmonresonance (SPR)
Fluorescence
Chemo-luminescence
White light interferometry
Magneticnanoparticles
Magneto-resistance(GMR, TMR)
Micro-nuclear magneticresonance
Colorimetric(lateral flow devices)
Quartz crystal microbalance
Surfaceacoustic waves
Amperometry
Voltammetry
Impedancespectroscopy Cantilevers
Optical MagneticElectro-
chemicalMassbased
Sample liquid
Signal readout& processing
Transducer
Receptor surface
Biosensor
Display of results
Waveguides Magneticnanoparticles
Funded by the European Union - GA 737089 7Kick-Off Meeting, Barcelona, Jan 17-18, 2017
αφ
Diagnostic biosensor development @ AIT
Funded by the European Union - GA 737089 8Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Photonic simulation cluster: 256 cores with 488GB RAM
1TB high performance SSD array and
5TB raid-5 HDD storage, automated backup
• 2D/3D finite difference time domain simulation packages
• variational mode matching tools for eigenmode calculation
• Floquet-Bloch solver for simulating waveguide grating couplers
Temperature controlled sample holder
and programmable fluid control system
with five fluid reservoirs
Fully equipped set-up for waveguide device characterization and biosensing experiments
3D FDTD simulation of photonic wire Bragg reflector
Photonic design & characterization
Funded by the European Union - GA 737089 9Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Mach Zehnder interferometer waveguide biosensor
binding events change phase
velocity of light in sensing arm
Concentration
Intensity
Biosensitive layer on top of waveguide provides selective binding and enrichment of biomolecules on sensor surface
substrate ns
core layer nwg
waveguide side view
sensing layer nslLight
intensity
H
wwaveguide front view
analyte
Funded by the European Union - GA 737089 10Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Human IgG detection in serum on optical waveguides
analyte & 2nd Ab
serum
analyte only
serum
Human IgG (hIgG) detection in serumLimit of detection with secondary antibodies:0.1nM (≙15ng/ml)
Funded by the European Union - GA 737089 11Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Miniaturization: optoelectronic integration
optoelectronic layerSi-photodiodes
silicon dioxid 1.5-2 µm
Si3N4 waveguidesfabricated with CMOS compatibleback-end process
input grating output grating
optical sensor element
silicon substrateTSV
contactpad
=850nm
Funded by the European Union - GA 737089 12Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Magnetic biosensors
Based on magnetoresistance (GMR, TMR)
Electronic readout: Stray field of magnetized markers alters resistance
magnetizing field
Si/SiO2
SiO2
APTES
sensor
Hex
Funded by the European Union - GA 737089 13Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Based on magnetoresistance (GMR, TMR)
Electronic readout: Stray field of magnetized markers alters resistance
0 10 20 30 40 50 600,00
0,05
0,10
0,15
0,20
0,25
0,30
GMR data
exp. fit function
GM
R [
%]
sensor surface coverage [%]
blank level
Magnetic biosensors
Funded by the European Union - GA 737089 14Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Based on magnetoresistance (GMR, TMR)
-300 -200 -100 0 100 200 300
11,20
11,25
11,30
11,35
11,40
11,45
11,50
11,55
11,60
11,65 parallel
perpendicular
to annealing direction
res
ista
nc
e (
Oh
m)
magnetic field (Oe)-300 -200 -100 0 100 200 300
11,20
11,25
11,30
11,35
11,40
11,45
11,50
11,55
11,60
11,65 parallel
perpendicular
to annealing direction
res
ista
nc
e (
Oh
m)
magnetic field (Oe)
-300 -200 -100 0 100 200 300
11,20
11,25
11,30
11,35
11,40
11,45
11,50
11,55
11,60
11,65 parallel
perpendicular
to annealing direction
resis
tan
ce (
Oh
m)
magnetic field (Oe)
2.5 nm MgO
Magnetic biosensors
Funded by the European Union - GA 737089 15Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Based on magnetoresistance (GMR, TMR)
Electronic readout: Stray field of magnetized markers alters resistance
Easily integrated, no background signal
Flexible: average signal across certain area ↔ single marker resolution
Example: CMOS-integrated TMR biochip (mnt-era.net project MinoLab)
Magnetic biosensors
Funded by the European Union - GA 737089 16Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Based on magnetoresistance (GMR, TMR)
Electronic readout: Stray field of magnetized markers alters resistance
Easily integrated, no background signal
Flexible: average signal across certain area ↔ single marker resolution
Example: CMOS-integrated TMR biochip (mnt-era.net project MinoLab)
Magnetic biosensors
Funded by the European Union - GA 737089 17Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Tunneling magnetoresistance (TMR) biosensor
Detection of individual magnetic beads by CMOS-integrated TMR sensing cells
10 µm
Wheatstone bridge sensor with magnetic beads at surface (red bold curve) shows clearly distinguishable characteristics to out-of-plane magnetizing fields
Results of mnt-era.net project MinoLab & FP7 project e-brains
-200 -150 -100 -50 0 50 100 150 200 250
-50
-40
-30
-20
-10
0
10
20
30
40
dig
ita
l o
utp
ut sig
na
l (o
ffse
t b
y a
ve
rag
e)
out-of-plane magnetizing field (Oe)
sensor with magneticbeads on top
München
Funded by the European Union - GA 737089 18Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Magnetic nanoparticle-based molecular biosensing
Mix & measure detection principle – homogeneous assay
+sample
cuvette
Funded by the European Union - GA 737089 19Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Analyte molecule
specifically binds to
functionalized nanoparticle
Magnetic nanoparticle-based molecular biosensing
Funded by the European Union - GA 737089 20Kick-Off Meeting, Barcelona, Jan 17-18, 2017
On analyte
binding, a
measureable
physical property
of the nanoparticle
changes
Magnetic nanoparticle-based molecular biosensing
Funded by the European Union - GA 737089 21Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Magnetic nanoparticle-based molecular biosensing
Funded by the European Union - GA 737089 22Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Magnetic nanoparticle-based molecular biosensing
Funded by the European Union - GA 737089 23Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Nanoprobes: noble-metal shell coated Co-nanorods
Co-nanorods synthesized by
organometallic approach
• Particles obtained from LPCNO-
INSA, Toulouse, France
• Single crystalline hcp Co
• Usually dissolved in organic
solvents with HDA as ligand
magnetic core length 40..200 nm
magnetic core Ø 6 nm
Nobel metal coating procedure
• Interlayer on Co-nanorods reduces
difference in surface energies
between Co and Pt/Au
• Overgrowth of Au on Pt-coated Co-
nanorods
Magnetic nanoparticle-based molecular biosensing
Funded by the European Union - GA 737089 24Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Diagnostic prototype instrument
-200 -100 0 100 200
2.18
2.22
2.26
2.30
2.34
time (ms)
-2
-1
0
1
2
φ
small coil driving voltage (V)
photodetectorsignal (V)
power supply unitMeasurement unit & notebook
laser diode
photodetector
coil pairs
PC audio amplifier
Lock-in amplifier
sample
Funded by the European Union - GA 737089 25Kick-Off Meeting, Barcelona, Jan 17-18, 2017
0 1 2 3 4 5 6 7 8
0
2
4
6
8
10
12
14
ph
ase
lag
diffe
ren
ce
(°)
sHER2 concentration (nM)
HER2 assay results
• Buffer: 10 mM HEPES, 150 mM NaCl, 0.05 v% Tween 20, pH 7.4• Nanoparticle concentration: 10 pM• Rotating magnetic field: 1 kHz frequency, 10 mT amplitude
analyte & 2nd Ab
10% serum
analyte only
buffer
analyte & 2nd Ab
10% saliva
analyte & 2nd Ab
buffer
Extrapolated limit of detection:2 nM in undiluted saliva
Funded by the European Union - GA 737089 26Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Electrochemical detection of proteins
Detection of the hepatitis antibody
Au surface
immobilized antigen
blocking
surface functionalizationNyquist plots for different hepatitis B antibody concentrations
Funded by the European Union - GA 737089 27Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Fully printed electrochemical sensors
Funded by the European Union - GA 737089 28Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Impedance Sensing on Adherent Cell Cultures
• Long-term monitoring for toxicological investigations• (General) Monitoring technique for cell research?
• Impedance measurement is a possible way• Wireless approach to give operators in a lab a maximum freedom of
movement and avoids error-prone cable connections
Funded by the European Union - GA 737089 29Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Impedance Sensing on Adherent Cell Cultures
3,5 h
Funded by the European Union - GA 737089 30Kick-Off Meeting, Barcelona, Jan 17-18, 2017
8 h
Impedance Sensing on Adherent Cell Cultures
Funded by the European Union - GA 737089 31Kick-Off Meeting, Barcelona, Jan 17-18, 2017
24 h
Impedance Sensing on Adherent Cell Cultures
Funded by the European Union - GA 737089 32Kick-Off Meeting, Barcelona, Jan 17-18, 2017
48 h
Impedance Sensing on Adherent Cell Cultures
Funded by the European Union - GA 737089 33Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Impedance Sensing on Adherent Cell Cultures
Adherent cells attach on bottom surface
Cell membrane with cytoplasm has higher impedance than surrounding (ionic conductive) culture media
Electrodes integrated in bottom surface
Cell adhesion
Settling Adhesion/Spreading
Funded by the European Union - GA 737089 34Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Impedance Sensing on Adherent Cell Cultures
Sensor Device Components
Funded by the European Union - GA 737089 35Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Impedance Sensing on Adherent Cell Cultures
Assembled sensor device
Funded by the European Union - GA 737089 36Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Impedance Sensing on Adherent Cell Cultures
RF-based system – Base station
Antenna coil
to PC
Wireless transferof Energy, Data,and Commands
Base station
µC
MUX
RF frequencygenerator
RFID receiver
Multiwell plate with tags and
attached sensors
Funded by the European Union - GA 737089 37Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Impedance Sensing on Adherent Cell Cultures
Sensor tags
Funded by the European Union - GA 737089 38Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Impedance Sensing on Adherent Cell Cultures
Overall system
Sensors on tags within a
common rack
Tags with whole measurement
circuitry
Reader with connection to data
acquizition unit (e.g. PC)
Wireless energy and data
transfer
Funded by the European Union - GA 737089 39Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Impedance Sensing on Adherent Cell Cultures
Conditions
Cell line 3T3 Clone A31 (Mouse
fibroblast)
Cell culture medium DMEM with
10% Fetal Calf Serum (FCS)
~ 300µl per sensor device
Measurement period of about 10
minutes
Funded by the European Union - GA 737089 40Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Impedance Sensing on Adherent Cell Cultures
0 24 48 72 t [h]-24
3 Cell samples
Blank
1.0
0.5
1.5
2.0
2.5
3.0
Re
{}
[a.u
.]Z
sen
s
Measurements on 3T3 fibroblasts
50000 cells per sensor („>“ 100% confluence)
Blanks
Cells
Seeding
Medium
exchange
Funded by the European Union - GA 737089 41Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Impedance Sensing on Adherent Cell Cultures
Measurements on hMSC
50000 cells per sensor, cell differentiation
1.5 3.0 t [d]1.0 2.0 2.50.50
-15
-5
0
Re{
} . 10
[m
]Z
sen
s
28
-10
0
D
Funded by the European Union - GA 737089 42Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Impedance Sensing on Adherent Cell Cultures
Funded by the European Union - GA 737089 43Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Impedance Sensing on Adherent Cell Cultures
3 4 7 t [d]2
0
10
15
5 61
5
0
Re{
} . 10
[m
]Z
sen
s
28
D
Measurements on hMSC
50000 cells per sensor, cell differentiation
Funded by the European Union - GA 737089 44Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Impedance Sensing on Adherent Cell Cultures
Funded by the European Union - GA 737089 45Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Impedance Sensing on Adherent Cell Cultures
There is urgent need for investigation cell wall characteristics by a quick method.
Funded by the European Union - GA 737089 46Kick-Off Meeting, Barcelona, Jan 17-18, 2017
Acknowledgments
Collaborators Funding organizations
NAMDIATREAM project
(EU NMP4-LA-2010-246479)
SPONSORED BY THE
Project # 823985
mnt-era.net project MinoLab
Funded by the European Union - GA 737089 47Kick-Off Meeting, Barcelona, Jan 17-18, 2017
www.ait.ac.at/profile/detail/Heer-Rudolf
Thanks for your attention.