lab on a chip integration - optolabcardslide 8 objective: to solve a social need real food, human,...
TRANSCRIPT
Slide 1
Lab on a Chip Integration
© Optolabcard
© Ikerlan
Slide 2
IntroductionDescription
State of the artObjectivesStrategy
Fabrication and packagingResults overview Exploitation potentialConclusions
Summary
Slide 3
www.ikerlan.es
Ikerlan is a private research centreThere are 230 researchers working in 12 groups
Slide 4
• Home: Domestic appliances (Fagor), elevators (Orona), locks (Azbe)• Spare time: Azkoyen coffee machines, vending machines• Travelling: Bilbao, Madrid and Hong-Kong underground (CAF), buses (Irizar)• Working: Fagor CNC and presses, Mecalux and Ulma warehouses• Sports: Orbea cycles, Dikar-Wingroup gyms tools
Introduction
Slide 5
http://www.cepheid.com/Sites/cepheid/content.cfm?id=165
CEPHEID SOLUTION
•Self Contained Cartridges for Integrated sample preparation - for automated and self-contained testing.
•They use fluorescence method but it is not portable
State of the artThe most remarkable available product
Slide 6
State of the art
SAMSUNG LTD.:
Jeong-Gun Lee, Kwang Ho Cheong, Nam Huh, Suhyeon Kim, Jeong-Woo Choi and Christopher Ko, Microchip-based one step DNA extraction and real-time PCR in one chamber for rapid pathogen identification, Lab Chip, 2006, 6, 886 - 895.
They report a novel method for efficient DNA extraction and subsequent real-time detection in a single cartridge by combining laser irradiation lysis and magnetic beads.
A remarkable research work
Slide 7
Dynabeads+Antibody+Bacteria On-chip Lysis and PCR
10.0010.5011.0011.5012.0012.5013.0013.5014.0014.5015.00
0 5 10 15 20 25 30 35 40
Time [s]
Phot
omul
tiplie
r Vo
ltage
[V]
Dynabeads+Antibody+Bacteria On-chip Lysis and PCR
10.0010.5011.0011.5012.0012.5013.0013.5014.0014.5015.00
0 5 10 15 20 25 30 35 40
Time [s]
Phot
omul
tiplie
r Vo
ltage
[V]
158 bp
CT=19
+ Lysis + qPCRReal sampleconcentration
State of the art
María Agirregabiria et al. Concentration, purification, lysis and real-time PCR on a single SU-8 microchamber for rapid detection of Salmonella spp. In faeces. The 11th International Conference on Miniaturized Systems for Chemistry and Life Sciences, microTAS 2007 Conference, 7-11 October, Paris ( France )
Another remarkable research work
Slide 8
Objective: to solve a social need
Real food, human, environmental sample
preparation
Integration of Sample preparation and
Detection
Available solutions
Current strategyLab on a Chip
strategy
Microtechnology+
Biology
+
High demandof point of
care devices 1. High throughput fabrication process
2. Few materials involved3. Authentic sample
preparation4. Simple fluidic control5. High sensitivity and
specificity6. Small amount of
measured sample7. Reagents stored within
the Lab on a Chip8. Fluidic microdevices
easy to handle9. Low cost disposable
components10. Regulatory issues
Restrictions
Jesus M.Ruano-Lopez. Fabrication strategies to integrate 3D microfluidic networks with biosensors to manufacture Lab on a Chip devices. Measurement + Control, Vol.40, No. 4, pp. 111-115
Slide 9
IkerlanIkerlan MicrosMicrosyystem stem Group StrategyGroup Strategy
PProductroduct DevelopmentDevelopment
Slide 10
Photo of a PCR chamber fabricated
on PyrexM.Agirregabiria et al. Fabrication of SU-8 multilayer
microstructures based on successive CMOS compatible adhesive bonding and releasing steps . Lab on a chip,
Vol. 5, No. 5, pp. 545-552F J Blanco et al. Microfluidic-optical integrated CMOS compatible devices for label-free biochemical sensing,
Journal of Micromechanics and Microengineering, Vol.16, No.5, pp. 1006-1016
Fabrication of Lab on Chips
Slide 11
Patented: Polymer microfluidic-electrical technology tofabricate flexible devices
© Ikerlan-IK4
Fabrication of flexible Lab on Chips
Slide 12
Other Polymeric microfluidic chipsHot embossing and Thermal bonding without channel collapsePolymers such as PMMA, PC and COC
M.T.Arroyo, L.J.Fernández, M.Agirregabiria, N.Ibañez, J.Aurrekoetxea and F.J.Blanco. Novel all-polymer microfluidicdevices monolithically integrated within metallic electrodes for SDS-CGE of proteins. Journal of Micromechanics and Microengineering, Vol. 17, No. 7, pp. 1289-1298.
Slide 13
PackagingAfter Kapton releasing, easy electric/fluidic connectionWithout glue/wires, easy device replacement
open/closeScrews
PCB
Top capsule
O-ringsPCR chip
Bottom capsule
External tube
External electriccontact
Spring loadedcontact
Slide 14
Multiple Calibration
3.080.86TCR
23.354.1R (Ω)
StD betweenwafers (%)
StD in thewafer (%)
Average R at 30ºC = 84.36Ω
Average TCR = 1979
∆T = 0.7ºC∆T = 2.53ºC
Slide 15
Sample concentration plus Real Time PCR on a LabonachipCZE and GCE electrophoresis of DNA and proteins within a flex devicePolymeric Microfluidic probesA hybrid integration example of a Lab on a Chip
Ikerlan Microsystem applications
Slide 16
Sample concentration plus Real Time PCR on a Lab on a Chip
Slide 17
Experimental methodology
Pyrex
Pyrex
Pyrex
NS
SN
Sample
Pyrex
NS
SN
PCR reagents
Bacteria concentration+ washing
Lysis
PCR mixture loading
qPCR
Magnetic bead
Bacteria
DNA
Slide 18
Dynabeads+Antibody+Bacteria On-chip Lysis and PCR
10.0010.5011.0011.5012.0012.5013.0013.5014.0014.5015.00
0 5 10 15 20 25 30 35
Time [s]
Phot
omul
tiplie
r V
olta
ge [V
]
Dynabeads+Antibody+Bacteria On-chip Lysis and PCR
10.0010.5011.0011.5012.0012.5013.0013.5014.0014.5015.00
0 5 10 15 20 25 30 35
Time [s]
Phot
omul
tiplie
r V
olta
ge [V
]
human faecal sample
Swab with real sample
bacteria + beads + antibodies
Salmonella DNA collection Sample concentration Lysis and real-time PCR ON CHIP
Magnet
human faecal sample
Cellsbeads
RNA collection
Lyisisbuffer
Sample concentration
Magnet
RT PCR OFF CHIP
RTPCR ON CHIP IN PROGRESS
PCR Lab on chip results
CT=19
Method and device patented
Slide 19
CZE and GCE electrophoresis of DNA
and proteins within a flex device
Separation of trypsin inhibitor and gliceraldehyde
2
2,5
3
3,5
4
4,5
0 50 100 150 200Separation time (sec)
Det
ecto
r sig
nal (
mV)
1
2
)
Separación de inhibidor de tripsina y gliceraldehído sin gel, a 343V/cm, a una distancia de 10 mm
6
7
8
9
10
11
12
13
14
15
0 50 100 150 200
Tiempo de separación (segundos)
Seña
l del
det
ecto
r (m
V)
4
6
8
10
12
14
0 20 40 60 80 100 120 140
Tiempo de migración (segundos)
Seña
l del
det
ecto
r (m
V) Visualización del amplificado del genotipo WT de 250 bp, en el dispositivo
miniaturizado
Maria Agirregabiria, Francisco Javier Blanco, Javier Berganzo, AsierFullaondo, Ana María Zubiaga, Kepa Mayora, Jesús Miguel Ruano-López. Sodium dodecyl sulfate-capillary gel electrophoresis of proteins in microchannels made of SU-8 films. Electrophoresis, Vol.27, No. 18, pp. 3627-3634 *In collaboration with The
Basque Country UniversityBiology Dept.
Slide 20
Amperometric detection of the electrophoresis1. Sample preparation by stacking
2. Integration of detection avoidingcomplex optical detectors
AmperometricSensor*
*In collaboration with Facultad de Química de la Universidad de Oviedo
-3
-1
1
3
5
7
9
11
13
0 15 30 45 60 75 90 105 120 135 150
tiempo (s)
Inte
nsid
ad (n
A)
+500V+750V+1000V+1250V
Vsep (V) ip (nA) tm (s) w1/2 (s)250 4.6 166.5 24500 5.8 76.8 8.7750 5.8 57 5.11000 6.9 46.2 4.51250 6.7 36.3 3.3
Condiciones: Viny = +250 V, tiny = 5 sEd = +0.5 V, Buffer: 50 mM Tris-Bórico pH = 7.0
Analito: Dopamina (DA) – 100 mM
Slide 21
An Instrument to identify extinct life on Mars
A hybrid integration example of a lab on a
Chip
*In collaboration with INTA-CAB and SENER
Slide 22
The Life Marker Device should be an instrument capable to identify evidence of extinct life on the exploration of Mars. The approach proposed on this document is based on a well known techniques, protein microrrays, measuring fluorescence.
An Instrument to identify extinct life on Mars
*In collaboration with INTA-CAB and SENER
Slide 23
20 µm
60 µm 20 µm
100 µm
Integratedelectrodes
Microfluidics
MultipleMicrofluidic channels
Ikerlan Microfluidicprobes
L.J.Fernández, M.Tijero, R.Vilares, J.Berganzo, K.Mayora andF.J.Blanco. SU-8 based microneedle for drug delivery in nanomedicine applications with integrated electrodes. The 11th International Conference on Miniaturized Systems for Chemistry and Life Sciences, microTAS 2007 Conference, 7-11 October, Paris (France).
Slide 24
Microfluidic probes results
0,0 0,2 0,4 0,6 0,8 1,0 1,20
10
20
30
40
50
60
70
80
90
100Channel dimensions (µm width x µm height)Red 200x25 Blue 150x25 Yellow 125x25Magenta 100x25Green 70x25Grey 50x25
Y=1,75+30,73*X;R=0,99
Y=1,35+41,56*X;R=0,99
Y=1,03+50,23*X;R=0,99
Y=6,6+60,6*X;R=0,98
Y=4,6+72,1*X;R=0,99
Flow
(µl/m
in)
Pressure (bar)
Y=9,0+79,8*X;R=0,98
PC adquisition
system
Flowmeter
DiferentialPressure Sensor
Microfilter (0.2 µm)
Liquidinjectionsystem
Microfluidic chip
Experimental set up
• Microfluidic-electrical flexible/semiflexible polymer
• Integration electrical and chemical functions into tissues.
• Chronic ImplantsHigh Biocompability
Polymer devices can fit to the tissues and deform their shapes as the organs deform
Slide 25
Aguja encapsulada
Dispositivo para experimentación biomédica
Microfluidic probes results
*In collaboration with CNM Barcelona
Patent pending
Slide 26
Product´s Roadmap of two technologies: Lamination and Embossing
20102006 2007 2008
Lam
inat
ion
Hot
em
boss
ing
PlatformR&D
Embossed Samples
Available Product Pasives
Plataform I+DSU8+Laminates
DeviceqPCR
qPCRCommersialisation
2009
Other devices: CE,µPCR,concentrator
Product´s Roadmap of two technologies: Lamination and Embossing
20102006 2007 2008
Lam
inat
ion
Hot
em
boss
ing
PlatformR&D
Embossed Samples
Available Product Pasives
Plataform I+DSU8+Laminates
Truly Labonchips
LOCCommercialisation
2009
Other devices:
Current situation
Exploitation potential
Slide 27
Exploitation plan based on Ikerlan Chips
A New Business Model*
• Similar market prices offer a dramatic increase of potential cash flow • Sales price decreasing from 2,5 to 0,7 €/Cm2.• Less dependency on productivity
*In collaboration with the Strategic Innovation Area at Ikerlan
Dry filmsOLED
LOC Labcard
Slide 28
ConclusionsThe developed fabrication technology proves to be an excellent tool
for rapid prototyping, as well as for mass production of low cost disposable Lab on Chip.
We have developed a set of Lab-on-a-Chips fulfilling the restrictions to be mass produced
A truly Lab on a Chip where bacteria concentration from human faecal sample, lysis and DNA amplification of Salmonella spp. were successfully carried out on a single chip.
The advantages of the miniaturisation in terms of time and cost in comparison with the conventional electrophoresis methods have been proved
It has demonstrated the potential of Ikerlan Probes
A Lab on a Card has been created by integrating a Lab on a chip in a Smartcard
Slide 29
Acknowledgements
Basque Government and the Provincial Council for its financial support under the ETORTEK program.
Optolabcard european project.
To all the Ikerlan groups involved in our activities and special thanks to all the microsystem team.
OPTOLABCARDwww.optolabcard.com
Slide 30
Thank you for your attention
It is time for questions
Jesus M. Ruano-López
Slide 31
Nerve stimulationPaceMakers
cochlearcatheters
Implants
MicroneedleScalpelProbes
Microsurgery
In Vivo
Point of careCapillary Electrophoresis
DNA deteccion PCRCell studies
Lab-on-a-Chip
ProteomicsGenómicsPharma
Drug screening
Microarrays
In Vitro
BIOMICROTECHNOLOGY
Lab on Chip state of the art
Slide 32
State of the art
• The 90´s was a decade of incredible microfabrication process developments and transducing mechanism characterisation.
• However, few of these processes were transfered successfully into real applicationsbecause the difficulty of integrating them reliably in a mass-production.
• Due to the maturity of the microtechnology, it is now the moment to prioritise miniaturisation, sample preparation and optical detection
Slide 33
Planarmicroelectrodearrays (pMEAS)
Slide 34
http://www.multichannelsystems.com/
http://www.med64.com/products/medprobe.html
http://www.plexoninc.com/
Planar microelectrode arrays (pMEAS)Vital organs and tissues
Bioelectrical phenomena
Electrical activityat cellular level
Long-term monitoring ofthe activity of cell cultures
electronic signal amplification
signal processing software & data logging
to keep cells alive
Commercially available systems
Polymeric and flexible bottom substrates made of SU-8 and PDMS
PDMS
SU8Ti/Au electrodes Flexible MEAS made ofPDMS
+ planar electrodes& SU8 passivation
SU8
SU8Ni electrodes Flexible MEAS made of SU8
+ 3D electrodes& SU8 passivation
Flexible chips with Ni electrodes
Slide 35
Wafers substrate
Dry films
ImmunoPCRRT-PCR
NASBALionprobes
Skin absorption
OLED
Gelification
LOC Labcard Labcard ReaderSyringe
www.smartbiophone.com
LOC
Current Bio-MEMS convergence
Wafer depencencyNo sample prep or limitedBulky equipmentExpensive for dispose
MEMS based onfilms
Molecular biology reactions and biological techniques that simplify the LOC (e.g. gelification)
Ikerlan MST group Bio-MEMS convergence
Band-Aid
Exploitation potential