microfluidics for electrical biochip technology...l.blohm et al., “rapid detection of different...
TRANSCRIPT
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8th Workshop Low Flows in Medical Technology, September 24th 2014
Fraunhofer Institute for Silicon Technology (ISIT)
Microfluidics for Electrical Biochip Technology
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as well as in the event of applications for property rights.
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Lars Blohm, Department „Biotechnical Microsystems“
Biotechnical Microsystems
Fraunhofer ISIT
� Continuous Enzyme Sensors
� Glucose and Lactate Monitoring
� Multi-Pore Membrane Chips
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as well as in the event of applications for property rights.
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� Multi-Pore Membrane Chips
� Bioprocess Analytics
� Electrical Biochip Arrays
� Protein and DNA Detection
� Chromatography On-Chip
� Small Molecule Detection
� Mass Flow Sensors
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Research and Production at one Location
Microelectronics and Microsystems Technology
Fraunhofer ISIT
� IC Technology and Power
Electronics
� Packaging Technology for
Microsystems and Microelectronics
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as well as in the event of applications for property rights.
© Fraunhofer
� Micro Electrical Mechanical System
(MEMS)
� IC Design
� Biotechnical Microsystems
� Quality and Reliability
of Electronic Assemblies
� Integrated Power Systems
Continuous monitoring of lactate in sweat
during exercise for sports medicine
Electronic detection of lactate (ELaN)
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as well as in the event of applications for property rights.
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Motivation for continuous monitoring of lactate in sweat
• Individual check of endurance performance
• Non-invasive measuring of lactate during exercise
• Recognition of anaerobic threshold for continuous training control
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as well as in the event of applications for property rights.
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• Bio-compatible disposable sensor system for monitoring
• Low production costs by printed electronics
• Evaluation of fitness status via Smartphone,
PC or internet portal
Concept of the project (ELaN)
Label / Plaster
Data evaluation:
Smartphone, Server, PC
Data transmission
NFC
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as well as in the event of applications for property rights.
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Lactate sensor chipMeasurement
electronics
Data preparation
Energy supply
MemoryDisplay
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Principle of the enzyme sensor
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as well as in the event of applications for property rights.
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• Diffusion of the analyte through the pore membran
• Enzymatic reaction (e.g. lactate – lactate oxidase)
• Amperometric detection of hydrogen peroxide
- G. Piechotta et al., “Novel micromachined silicon sensor for continuous glucose monitoring”, Biosensors and Bioelectr. 21 (2005) 802–808
Layout and dimensions of the current lactate chips
• Chip size: 6 mm x 7 mm
• Pore membrane: 0,65 mm x 0,65 mm
• Pore size: 5 µm or 10 µm
• Amount of pores: 9, 25, 100
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as well as in the event of applications for property rights.
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Layout and dimensions of the current lactate chips
Counter electrode
Working electrodes
Reference elcectrode
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Temperature sensor
Conductivity electrodes
Heating resistor
Contact pads
Lactate chip and pore membrane with 100 Pores (ᴓ10µm)
← 650 µm →
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as well as in the event of applications for property rights.
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← 650 µm →
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Concentration series with lactate chip in a flow through cell
• Differential measurements for compensation
of interfering substances
• Flow rates ~ 50µl / min.
100
120
140
40 mM
50 mM
30 mM
Lactate chip with 25 pores (5 µm diameter)
Pt-working electrode: 500 mV in relation to IrOx-reference electrode
cu
rren
t [n
A]
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as well as in the event of applications for property rights.
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0 100 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400
0
20
40
60
80
reference signal (cavity without enzyme)
30 mM
20 mM
5 mM
10 mM
1 mM
cu
rren
t [n
A]
time [s]
0 mM
Sweat collection by microfluidic cartridge
FTS-105-01-X-
DV
ASIC (SO16)
NFC (SO28) CR2032
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as well as in the event of applications for property rights.
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CLP-105-02-X-D
Sweat collector
prototype
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Comparison of different measuring methods in an Ergometer test
• Increase in steps of 50 W
• Interval 3 min.
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as well as in the event of applications for property rights.
© Fraunhofer
- C.G.J. Schabmueller et al., « Micromachined sensor for lactate monitoring in saliva”, Biosensors and Bioelectronics 21 (2006) 1770–1776
∆ blood: photometer ▲ saliva: photometer • saliva: silicon sensor
Fraunhofer Lighthouse Project
(IBMT, IGB, IME, IPA, IPK, ISiT, ISI, IZM)
Cell-Free Bioproduction
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as well as in the event of applications for property rights.
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Cell-free protein production with integrated energy supply
• Scheme of pore membrane for ATP synthesis
• ATP-Synthase is just active in lipid double layer
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as well as in the event of applications for property rights.
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Micro-pores in silicon membranes and integrated micro electrodes
Chips with 4 and 16 pores on a Wafer Membrane with 16 pores / electrodes
--- 500 µm ---
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as well as in the event of applications for property rights.
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Membrane (1.8 µm thick)
on the bottom of a cavity
--- 500 µm ---
Pore (ᴓ 4 µm)
Electrode (ᴓ 50 µm)
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20121207 16-Po r e n -Go ld ch ip -PBS03n Ze lle _01.m p r
|Z |, log s pac ing v s . f req, log s pac ing Phas e(Z) v s . f req, log s pac ing #
|Z|/
Oh
m,
log
sp
ac
ing
100.000
-15
-20
-25
Technical membrane with16 Pores on a silicon chip
• Impedimetric detection of lipid double layer
• Electrochemical impedance spectroscopy
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as well as in the event of applications for property rights.
© Fraunhofer
f r e q / H z , l o g s p a c i n g
1 10 100 1.000 10.000 100.000
|Z|/
Oh
m,
log
sp
ac
ing
10.000
100.000 Ph
as
e(Z
)/de
g
-30
-35
-40
-45
-50
-55
20140624 TEOS-Chip-4Pore n+DiphPC-Grube -vorne Warne rze lle 2+Alubox-Schirm ung 10m V 33We rte -Se k -ohne Filte r-AgCl-PBS_01.m pr
I vs. time
I/p
A
110.000
100.000
90.000
80.000
70.000
60.000
50.000
Second brushing
of lipid in solvent
Lipid layer
on 1. pore
Lipid layer
on 2. pore
First brushing
of lipid in solvent
Formation of lipid bilayer on membrane chips with 4 pores
20140617 TEOS-Chip-4Pore n+DiphPC+PC-Laye r -Gr ube -vor ne War ne rz e lle 2+Alubox-Schir m ung 10m V 33Wer te -Se k -ohne Filte r-AgCl-PBS_01.m pr
I vs . time
30
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as well as in the event of applications for property rights.
© Fraunhofer
t im e /s
350300250200150100500
50.000
40.000
30.000
20.000
10.000
0
of lipid in solvent
Lipid layer
on 4. pore
Lipid layer
on 3. pore
t im e /h
2015105
I/p
A
28
26
24
22
20
18
16
14
12
10
8
6
4
2
0
-2
10 mV; ~0.1 pA => > 100 GΩ
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pH-measurement in bioreactors for cell-free bio production
• pH-sensitive iridium oxide microelectrodes
• Integration of the sensor chips in microfluidic bioreactors
• Measuring of proton gradient on micro pore membranes
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© Fraunhofer
400.000100.00025.0006.400
1.600 400 100 25
4002000
Ew
e/V
0,22
0,21
0,2
0,19
0,18
0,17
0,16
0,15
400.000 µm²
Evaluation of IrOx-microelectrodes for pH-measurement
on micro pore membranes
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as well as in the event of applications for property rights.
© Fraunhofer
t im e /s
4002000
Ew
e/V
0,28
0,27
0,26
0,25
0,24
0,23
0,22
0,21
0,2
t im e /s
100 µm²
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Biotechnische Mikrosysteme
Electrical Array-Biochips
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as well as in the event of applications for property rights.
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Technology of the electrical array biochips
Principle:
Sandwich-ELISA on gold electrode arrays.
Detection via single electrode redox-cycling.
array positions
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size: 8 x 10 mm²
array positions
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Position specific dispensing of biological components
spotter tips
25 droplets
of 400 pl
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passivation layer
(hydrophobic)
gold
(hydrophilic)
position 1 position 2
200 µm silicon350 µm 350 µm
silicon dioxide
Piezoelectric nanodispensing device
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Biochip spottet with different capture molecules
1. Adding a diluted serum or
whole blood sample
2. Adding the enzyme conjugate
3. Adding the substrate and
Immunoassay on an electrical biochip
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current signal
redox cycling
3. Adding the substrate and
electrical read out
Pump and flow rates for assay procedure
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Hepatitis-C (HCV) detection with the electrical biochip system
negative sample3000 positive sample3000
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as well as in the event of applications for property rights.
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negative sample
Core
NS3
NS4A
positive
control
negative
control
1 2 3 4 5 6 7 8 9 10 11 120
200
400
600
800
1000
1200
1400
1600
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2000
2200
2400
2600
2800
3000
slo
pe [nA
/min
]
positive sample
Core
NS3
NS4A
positive
control
negative
control
1 2 3 4 5 6 7 8 9 10 11 120
200
400
600
800
1000
1200
1400
1600
1800
2000
2200
2400
2600
2800
3000
slo
pe [nA
/min
]
assay time:
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Microsystem for mobile analysis of biochemical molecules
“MEMS-Chromatography”
Biotechnical Microsystems
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Development of a portable analytical system on the basis of
liquid chromatographic separation processes
� Separation column with very large surface (porous material)
� Compatible with silicon technology
� Components for detection of biochemical substances
� Electrochemical detection methods
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� Electrochemical detection methods
� Integration of 3D-Column structures
and detection in a microsystem
� Process- and module integration
on wafer level
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3D-MEMS process for porous separation column
top view
column structure
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cross section
column structure
Column chip and test cartridge
column chip
detection chip
Integrated column
flow through cell
fluidic fittings
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as well as in the event of applications for property rights.
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Electrochemical detection of three different antibiotics with a
commercial column and gold microelectrodes
Au electrodes 600mV vs. IrOx reference
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as well as in the event of applications for property rights.
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Micro Electrical Mechanical Systems (MEMS)
„Mass Flow Sensors”
Microsystem Technology
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Temperature compensated Hot-Wire Mass Flow sensor with
recognition of direction
• Suitability
• for Gases and Fluids
• Recognition of
Flow Direction
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• Fields of Application
• Automotive
• Public Water Supply
• Medical Technology
Response function of flow rate
in volume per minute
• Wide dynamic range
• Independence of T (ambient)
Flow Sensor Concepts
• 2 Heaters
• 2 Temperature Sensors
Principle and test of the mass flow sensor
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Acknowledgments
Thanks to all involved project partner and colleagues from ISIT.
Department „Biotechnical Microsystems“Eric Nebling, Gundula Piechotta, Jörg Albers, Denise Rühmann, Simone Holz
Department „Microsystem Technology“Lutz-Martin Buchmann, Thomas Lisec, Peter Lange
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Thank you for your attention!