Microfluidics for Electrical Biochip Technology...L.Blohm et al., “Rapid detection of different...

8th Workshop Low Flows in Medical Technology, September 24th 2014

Fraunhofer Institute for Silicon Technology (ISIT)

Microfluidics for Electrical Biochip Technology

All rights reserved, also regarding any disposal, exploitation, reproduction, editing, distribution,

as well as in the event of applications for property rights.

© Fraunhofer

Lars Blohm, Department „Biotechnical Microsystems“

[email protected]

Biotechnical Microsystems

Fraunhofer ISIT

� Continuous Enzyme Sensors

� Glucose and Lactate Monitoring

� Multi-Pore Membrane Chips



© Fraunhofer

� Multi-Pore Membrane Chips

� Bioprocess Analytics

� Electrical Biochip Arrays

� Protein and DNA Detection

� Chromatography On-Chip

� Small Molecule Detection

� Mass Flow Sensors

Research and Production at one Location

Microelectronics and Microsystems Technology

Fraunhofer ISIT

� IC Technology and Power

Electronics

� Packaging Technology for

Microsystems and Microelectronics



© 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)



© Fraunhofer

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



© Fraunhofer

• 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



© Fraunhofer

Lactate sensor chipMeasurement

electronics

Data preparation

Energy supply

MemoryDisplay

Principle of the enzyme sensor



© Fraunhofer

• 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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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 →



© Fraunhofer

← 650 µm →

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]



© Fraunhofer

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



© Fraunhofer

CLP-105-02-X-D

Sweat collector

prototype

Comparison of different measuring methods in an Ergometer test

• Increase in steps of 50 W

• Interval 3 min.



© 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



© Fraunhofer

Cell-free protein production with integrated energy supply

• Scheme of pore membrane for ATP synthesis

• ATP-Synthase is just active in lipid double layer



© Fraunhofer

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 ---



© Fraunhofer

Membrane (1.8 µm thick)

on the bottom of a cavity

--- 500 µm ---

Pore (ᴓ 4 µm)

Electrode (ᴓ 50 µm)

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



© 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



© 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

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



© 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



© 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

Biotechnische Mikrosysteme

Electrical Array-Biochips



© Fraunhofer

Technology of the electrical array biochips

Principle:

Sandwich-ELISA on gold electrode arrays.

Detection via single electrode redox-cycling.

array positions



© Fraunhofer

size: 8 x 10 mm²

array positions

Position specific dispensing of biological components

spotter tips

25 droplets

of 400 pl



© Fraunhofer

passivation layer

(hydrophobic)

gold

(hydrophilic)

position 1 position 2

200 µm silicon350 µm 350 µm

silicon dioxide

Piezoelectric nanodispensing device



© Fraunhofer

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



© Fraunhofer

Hepatitis-C (HCV) detection with the electrical biochip system

negative sample3000 positive sample3000



© Fraunhofer

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

1800

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:

Microsystem for mobile analysis of biochemical molecules

“MEMS-Chromatography”

Biotechnical Microsystems



© Fraunhofer

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



© Fraunhofer

� Electrochemical detection methods

� Integration of 3D-Column structures

and detection in a microsystem

� Process- and module integration

on wafer level

3D-MEMS process for porous separation column

top view

column structure



© Fraunhofer

cross section

column structure

Column chip and test cartridge

column chip

detection chip

Integrated column

flow through cell

fluidic fittings



© Fraunhofer

Electrochemical detection of three different antibiotics with a

commercial column and gold microelectrodes

Au electrodes 600mV vs. IrOx reference



© Fraunhofer

Micro Electrical Mechanical Systems (MEMS)

„Mass Flow Sensors”

Microsystem Technology



© Fraunhofer

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



© Fraunhofer

Thank you for your attention!

Microfluidics for Electrical Biochip Technology...L.Blohm et al., “Rapid detection of different...

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