interfacing electrochemistry and dna interfacing … · 2011. 7. 12. · (chip substrate) n ss-dna...
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Groupe CREAB (Chimie de la Reconnaissance et Groupe CREAB (Chimie de la Reconnaissance et EtudeEtude des Assemblages Biologiques)des Assemblages Biologiques)DRFMC/SI3M, CEA de Grenoble, 17 Rue des Martyrs 38054 Grenoble DRFMC/SI3M, CEA de Grenoble, 17 Rue des Martyrs 38054 Grenoble cedex 9, Francecedex 9, France
PascalPascal MAILLEYMAILLEY:: [email protected] , [email protected]
℡℡ +33.(0)4.38.78.44.38
Interfacing electrochemistry and DNA for the design of genosensors
Interfacing electrochemistry and DNA Interfacing electrochemistry and DNA for the design of genosensorsfor the design of genosensors
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DNA Biochips PrincipleDNA Biochips Principle
ODN probesODN probes TransductionTransductiont
VHzI...
Signal
Hybridisation
t
VHzI...
Signal
Hybridisation
ImmobilisationImmobilisationAdsorptionAdsorptionChemisorptionChemisorptionCovalent GraftingCovalent GraftingEntrapment…Entrapment…
LocalisationLocalisationMicrofluidicMicrofluidic
HybridisationHybridisation
t
VHzI...
Signal
Hybridisation
DetectionDetection
Main goalsImmobilisationImmobilisation
TransductionTransductionhν hν’
Reference method : Fluorescence detection
Fluorophore streptavidinconjugate
Biotynilated target
hν hν’
Reference method : Fluorescence detection
Fluorophore streptavidinconjugate
Biotynilated target
F
F
n A-
n
F
F
n A-
n
F
F
n A-
n
Fluorescence response in a grey level scale
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1-Electrochemical Immobilisation
Using Polypyrrole Matrices
1-Electrochemical ImmobilisationElectrochemical Immobilisation
Using Polypyrrole MatricesUsing Polypyrrole Matrices
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ImmobilisationImmobilisation
DNA probe accessibility
Probe Integrity
Recognition properties
Recognition specificity/selectivity
Suitable with detection/transduction
11-- Electrochemical DNA Immobilisation using pyrrole and derivativeElectrochemical DNA Immobilisation using pyrrole and derivatives s
-n (2+δ) e -
-2nH+
+ nδA-n
,n A-
+
+06-0.8V vs. SCE
F
F
n A-
n
Mild conditions of immobilisation
Adressable
Routes of functionnalisation
Control of the polymer film coating
Physico-chemical activity of the matrixOther conducting polymers: PANIPANI, , PEDOTPEDOT, , PThPTh, , PJUGPJUG
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Electrode
NH
NH
NH
+
Charge p-
Electropolymérisation
-n x (2.3 e-, -2H+)
n
1.11.1-- DNA Immobilisation through electrostatic interactions with PPyDNA Immobilisation through electrostatic interactions with PPy
Polypyrrole doppingPolypyrrole dopping J . Wang et al, Langmuir 16 (2000) 2269G.G. Wallace et al, Synth. Met. 123 (2001) 279A. kumar et al, Chem. Commun (2004) 820 [PEDOT]
DNADNA CounterionCounterion
Electrostatic interactionsElectrostatic interactions D.S. Minehan et al, J. Macromol. Sci. A (Pure Appl. Chem.) 38 (2001) 1245
EEpp EEoxox
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1.21.2-- Immobilisation by postImmobilisation by post--functionnalisation of PPy matricesfunctionnalisation of PPy matricesBy covalent graftingBy covalent grafting H. Korri-Youssoufi et al, J. Am. Chem. Soc. 119 (1997) 7388
T.-Y Lee et al, Anal. Chem. 73 (2004) 5629 [PTh]
NH
OOH
NH
OO N
O
O
0,6 0,4
n
NH
OOH
NH
OO N
0,6 0,4
n
HCCT-AAG-AGG-GAG-TG
NH
OOH
NH
OO N
O
O
0,6 + 0,4
Electropolymerisation DNA grafting
By complexationBy complexationJ. Janata et al, J. Am. Chem. Soc. 125 (2003) 324
EEpp
By biological anchoringBy biological anchoringA. Dupont et al, Anal. Chim. Acta. 515 (2004) 271
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pyrrole : intelligent glue for biological applications
ELECTROCHEMISTRY BIOLOGY OPTICS
(applications) (detection)
CHEMITRY (chip substrate)
N
ssss--DNADNA
PeptideBiotinBiotin--pyrrolepyrrole
StreptavidinStreptavidin--pyrrolepyrrole
pyrrolepyrrole tooltool--boxboxds- DNA
1.31.3-- Electrodeposition and biochips : the PPy toolElectrodeposition and biochips : the PPy tool--boxbox
Protein
Sugars
Microorganism
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1.3.11.3.1-- DNA direct grafting on functionnal substrates: MICAMDNA direct grafting on functionnal substrates: MICAM™™
Livache al., Nucleic Acid Res., 22 (1994) 2915
O
O
P O-
O
O
ODN
HO O
NH
N
N
NH
(CH 2 )2
N
CH3
H
N
H
N
O
O
P O-
O
O
ODN
HO O
NH
N
N
NH
(CH 2 )2
N
CH3
H
N
H
N
H
N
1.1. ODNODN--derivatized pyrrolederivatized pyrrole
2.2. Multiplexed electrodes arrayMultiplexed electrodes array
3.3. ODN immobilisation ODN immobilisation viavia electrochemical addressingelectrochemical addressing
4.4. Fluorescence detectionFluorescence detection
Electrode 1 Electrode 2 Electrode 3
Electrode 1 Electrode 2 Electrode 3
EpEp
EpEp
Puce ADN ∼ MICAM® 128 dots (diam. = 50µm)Puce ADN ∼ MICAM®Puce ADN ∼ MICAM® 128 dots (diam. = 50µm)128 dots (diam. = 50µm)
Hepatitis C Virus genotyping”Livache et al., Anal.Biochem.,
255 (1998) 188
24 mM
2 to 40 µM
MMultiplexed IIntegrated CChip for AAdvanced MMicroanalysis
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1.3.21.3.2-- Wafer levelWafer level MICAMMICAM™™ processprocess
CMOS MUX CHIP 1 CMOS MUX CHIP 2 CMOS MUX CHIP N
Counter / Reference Electrode
Electrolyte
Oligo-Pyrrole Pyrrole
Active CMOS 100 mm wafer
Electronic boardPotentiostatComputer
From P. From P. CaillatCaillat, CEA/LETI, CEA/LETI
Electro-copolymerisation onultra thin electrolyte layer
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1.3.31.3.3-- DNA direct immobilisation on bulk substratesDNA direct immobilisation on bulk substrates
µ-Counter Electrode
EpEpBioactive PPyBioactive PPy SpotSpot
µ-Counter Electrode
EpEpBioactive PPyBioactive PPy SpotSpot
Localisation of the electrical field
SECMSECM
Miniaturised electrochemical cell
ElectrospottingElectrospotting
z
D
A
B
C
plat inum counter electrode
pipette t ippo lypy rrole spots
x
y
CECE WEWEPotentiostat
z
D
A
B
C
plat inum counter electrodeplat inum counter electrode
pipette t ippo lypy rrole spots
x
y
CECE WEWEPotentiostat
How to fabricate a multiparametric sensor ?
How to structurate the bulk and/or optical surfaces ?
Guedon et al, Anal. Chem., 72 (2000) 6003
Evans et al, Submited to Electrochem. Commun.Fortin et al, Electroanalysis in press
240 µm
teflon
Cavity of Inox needle
240 µm240 µm
teflon
Cavity of Inox needle
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1.3.31.3.3-- Electrospotting : Principle and films characteristicsElectrospotting : Principle and films characteristics
-2
-1.5
-1
-0.5
0
0.5
1
1.5
2
-2 -1 0 1 2 3 4
Voltage pulse (V)
Wor
kin
g E
lect
rod
e
Pot
enti
al (
V/S
CE
)
t1 t1+xInput voltage-impulsion
t1 t1+xTime(msec)
Up= Ui+∆U
U(V)
Ui - Uf
∆tUp= Ui+∆U
U(V)
Ui - Uf
∆t
Time(msec)
Up= Ui+∆U
U(V)
Ui - Uf
∆tUp= Ui+∆U
U(V)
Ui - Uf
∆t
Input voltage-impulsion
Generation of nm thick Ppy films
2D like biological structures
Compatibility Compatibility with SPRi with SPRi transductiontransduction
Reproducibility of the fabrication process
Homogeneity of the probes repartition
Specificity of the recognition event
Spot size (800 µm 250 µm)
Robustness-reusability of the multisensor
49
50
51
52
53
Arb
itrar
y un
its
2200 2400 2600 2800 3000 3200 3400 3600 3800
Time (s)
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1.3.31.3.3-- SubstrateSubstrate structuration structuration using using SECMSECM
N-(CH2)3-(O-CH2-CH 2)2-O-(CH 2)3-NH-CO-(CH2)5-NH-CO-(CH 2)4S
HN NH
O
Biotinylated-pyrrole monomer
-0.04
-0.02
0
0.02
0.04
0.06
0.08
0 2 4 6 8
Temps (s)
Co
ura
nt
(µA
)
0
0.2
0.4
0.6
0.8
Po
tentiel (V
/EC
S)
G O x E
GlucoseH2O2O2
e-
xy
z
G O x E
GlucoseH2O2O2
e-
xy
z
µ-Counter Electrode
EpEpBioactive PPyBioactive PPy SpotSpot
µ-Counter Electrode
EpEpBioactive PPyBioactive PPy SpotSpot
Detection of the biologically active spot using the
generation /collection mode
0 50 100 150 2000
50
100
150
200 i / nA
0.400.440.480.520.560.600.640.680.720.760.800.840.880.920.961.001.041.081.121.161.20
X Axis / µm
Y A
xis
/ µm
0 2 0 0 4 0 0 6 0 0 8 0 0 1 0 0 0
0 . 2
0 . 3
0 . 4
0 . 5
0 . 6
0 . 7
i / n
A
Y Ax is d isp lacement / µm
PBS 0.1 M pH 7[glucose] 10 mMEtip = 0.6 V/SCESpeed 7.5 µm s- 1
Validity of the conceptSSpot size reduction compared to spotting
Application to DNA spots generation
SECM detection ?
200 µmParameters:Tip diameter and shapeDistance tip-substrateDeposition time (100-500 ms)Deposition potentialMonomers concentrations
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2-Hybridization detection General considerations
2-Hybridization detection Hybridization detection General considerationsGeneral considerations
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2.12.1-- What’s the signal ? What’s the signal ?
Substrat
Couche biosensible
Substrat
Couche biosensible
Molecular recognition
No metabolic activity
What has changedWhat has changed
How to labell the recognition eventHow to labell the recognition event
Matrix
or
Labell
QCM
SPR
Photocurrent…
Real time Real time TransductionTransduction
Fluorescence
Redox intercalators
Microparticles
Enzymes
DNA electroactivity
Impedance
Conductivity
End point End point detectiondetection
IndirectIndirectDirectDirectSignal
t
Signal
t
Signal
t
Signal
LabellingLabellingAccumulationAccumulationAmplificationAmplification
e-?
∆∆E, i, Z, Y, σσ
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2.22.2-- Properties of the recognition signals Properties of the recognition signals
Direct or labelled reading
Detection or transduction (kinetic approach)
Positive or negative signal (dynamic range)
Parrallel or one-point (imaging)
Amplification (limit of detection)
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3-Electrochemical detection
of DNA hybridization:The conducting polymer approach
3-Electrochemical detection Electrochemical detection
of DNA hybridization:of DNA hybridization:The conducting polymer approachThe conducting polymer approach
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3.13.1-- PhysicoPhysico--chemical behaviour of PPy and other conducting polymerschemical behaviour of PPy and other conducting polymers
-0,16 0 0,16 0,32 0,48
-0,16
-0,12
-0,08
-0,04
0
0,04
0,08
0,12
0,16
0,2
0,24
I (m
A c
m-2
)
E (V/ECS)
1
2nnδδ AA--nnδδ AA-- --nnδδee----nnδδee--
, δ, δ AA--, δ, δ AA--
RedRed OxOx
nnN
H nnN
H nn
δδ++N
H nn
δδ++N
H
δδ++N
H
Semi-condutor (p) Conductive (bipolaron)
BC
BV
Eg
BC
BV
σσ S cm-1
- 106
-104
-102
-1
-10-2
-10-4
-10-6
-10-8
-10-10
-10-12
-10-14
-10-16
Métaux
Semi-conducteur
Isolant
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3.23.2-- Chronoamperometric hybridization detection at ODNChronoamperometric hybridization detection at ODN--doped Ppy filmsdoped Ppy films
Probe : (dG) 20
J . Wang et al, Anal. Chim. Acta 402 (1997) 7
Comp (dC)20
Non-comp
(dA)20 or (dT)20
dA20dT20
dC20 dC20dC20
(dG) 20 probe
(dA) 20 probe
-
+
SignalSignalReversible transient signalChange in film capacitance or conductivityCurrent until charge compensation
Comp : volume effectNon-Comp: adsorption
+0.15 V vs. Ag/AgCl
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A. kumar et al, Chem. Commun (2004) 820
3.33.3-- PEDOT conductivity as probing toolPEDOT conductivity as probing tool
Probe
Comp
Non-comp
Nanopore membraneGold thin film
ss-DNA doped PEDOT
∆∆∆∆R/RR/R00
SignalSignalDecrease of the PEDOT intrinsec conductivityHindrance of ionic transferEnhanced resistance variation with the duplex lenghtNo dependance of σ on the electrolyteDynamic range : 10-8 to 10-5 g mL-1
Potential (V)
n
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NH
OOH
NH
OO N
0,6 0,4n
HCCT-AAG-AGG- GAG-TG
H. Korri-Youssoufi et alJ. Am. Chem. Soc. 119 1997 7388
3.43.4-- PPy electroactivity as transduction vectorPPy electroactivity as transduction vector
Comp
Non-comp
a)a) NativeNativeb), Nonb), Non--complementarycomplementary
c) to e) Complementaryc) to e) Complementary10µM to 0.1 10µM to 0.1 mMmM
nnδδ AA --nnδδ AA -- --nnδδee----nnδδee--
, δ, δ AA --, δ, δ AA --
RedRed OxOx
nnN
H nnN
H nn
δδ++N
H nn
δδ++N
H
δδ++N
H
AA--
AA--
J. Janata et al,J. Am. Chem. Soc. 125 (2003) 324
Hindrance of the doping (ion exchange) processHindrance of the doping (ion exchange) process DNA hybridisation transistor
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3.53.5-- Redox probe functionnalised polymer as transduction vectorRedox probe functionnalised polymer as transduction vector
ODNODNprobeprobe
Detection limit 10 Detection limit 10 nMnM
H. Korri-Youssoufi et alAnal. Chim. Acta 469 (200)2 85
M.C. Pham et al, Anal. Chem. 75 (2003) 2478
Comp
Non-comp SignalSignalAccessibility of the redox sitesEnhanced cation exchange
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3.63.6-- Impedance spectroscopy of polymerImpedance spectroscopy of polymer--DNA filmsDNA films
Tae-Young Lee et al, Anal. Chem. 73 (2001) 5629
Hybridization
Comp
∆∆∆∆ZZ
SignalSignalDecrease of the electrochemical impedanceDependance on film thickness (and morphology)Discrimination of point-mutations
OCPOCP
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Photocurrent
3.73.7-- Photoelectrochemical behaviour of PPy as tansduction mediatorPhotoelectrochemical behaviour of PPy as tansduction mediator
O
O
P O-
O
O
ODN
HO O
NH
N
N
NH
(CH 2 ) 2
N
CH 3
n
H
N
m
H
N
O
O
P O-
O
O
ODN
HO O
NH
N
N
NH
(CH 2 ) 2
N
CH 3
n
H
N
m
H
N
H
N
VB
CBe-
hí
Lumière
Noir
PPyPPy Semiconductor in the neutral reduced stateSemiconductor in the neutral reduced state
Lassalle et al, J. Electroanal. Chem.509 (2001) 48
0,04
0,08
0,12
0,16
0,2
1 2 3 4 5Energie(eV)
0
Copol+ODNc
3,65
Copol
Copol+ODNnc
0,04
0,08
0,12
0,16
0,2
1 2 3 4 5
Energie (eV)
0
Copol+ODNc
3,65
Copol
Copol+ODNnc
0 2 4 6 8 10 12 14 16 18 20
0,68
0,72
0,76
0,8
0,84
0,88
0,92
0,96
1
Temps (min)
iph
(t)
/ ip
h(0
)
0 2 4 6 8 10 12 14 16 18 20
0,68
0,72
0,76
0,8
0,84
0,88
0,92
0,96
1
Temps (min)
iph
(t)
/ ip
h(0
)
i (m
A W
-1)
Dark
Light
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4-Electrochemical detection
of DNA hybridization at PPy-DNA films using
Redox labells
4-Electrochemical detection Electrochemical detection
of DNA hybridization at of DNA hybridization at PPyPPy--DNA films usingDNA films using
Redox labellsRedox labells
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4.14.1-- Redox molecules in interaction with dsRedox molecules in interaction with ds--DNADNA
Minor groove Minor groove bindersbinders
InteractionsInteractions
IntercalatorIntercalatorππππ StackingStacking
Threading Threading agentagent
Hoecsht 33258
Daunomycin
S. Takenaka, DNA and RNA binders, M. Demeunynck et al Eds, Wiley-VCH (2002) 225-246
Metal complexMetal complex
ElectrostaticIntercalation
Minor Groove
S Takenaka et al , Anal Chem72 (2000) 1334
J. Barton et al, Nucl.Acid Res 27 (2000) 4830
Methylene blue
M. Mascini et al, Fresenius J.. Anal. Chem. 367 (2000) 364.
Wang et al, Anal. Chem., 68 (1996) 2629
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N
N
O
A B C
D
1
2
3
4
56
7
8
9
1011
8H-8oxo-pyrido[4,3,2-mn]acridine
N
N
Cl
O
SOH
COB 137COB 137
N
N
Cl
O
HN
NHH2N
COB 142COB 142
-3.5
-2.5
-1.5
-0.5
0.5
1.5
2.5
-600 -300 0 300
E (mV)
i ( µµ
A)
-15-10-505
101520
-700 -400 -100 200
E (mV)
i (µµ
A)
-30
-20
-10
0
10
20
30
-600 -300 0 300
E/V vs. Ag/AgCl
I/µA
R2 = 0.9974
0
10
20
30
4 8 12 16
n1/2
/v1/2
s-1/2
I/µ
A
N
N
Cl
O
HN N
COB 136COB 136
4.1.14.1.1-- PyridoPyrido--acridones : a new family of redox intercalatorsacridones : a new family of redox intercalators
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Elaboration du capteur
Film Polypyrrole
Adsorption d’ADN(SS ou DS
Interaction avec Intercalant
EEDétection
Elaboration du capteur
Film Polypyrrole
Adsorption d’ADN(SS ou DS
Interaction avec Intercalant
EEDétection
Electrochemical detection of ds-DNA adsorbed at PPy
0
1
2
3
4
5
-400 -300 -200 -100 0
E/V vs. Ag/AgCl
I/nACOB 136 in solution
With ss Calf Thymus DNAWith ds Salmon DNA
0
0.5
1
1.5
2
2.5
0 50 100 150
Time/min
I/µA
0
1
2
3
4
-350 -250 -150
E/V vs. Ag/AgCl
I/µA
Slow kinetic of the intercalation processSelectivity of the recognition signal
Strong interaction with ds-DNANon-specific interactions with ss-DNA
DNA/COB 136 interactions in solution
4.1.24.1.2-- Electrochemical approach of COB136/DNA interactionsElectrochemical approach of COB136/DNA interactions
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ProbeProbePyr T5 5’AG AGT GCC TTG ACG ATA
CAG CTA ATT CAG AAT CAT TTT GT 3’
TargetsTargets10c10c 5’-bio-AA GGC ACT CT3’
20c20c 5’-bio-CTG TAT CGT CAA GGC ACT CT3’
40c40c 5’-bio-AC AAA ATG ATT CTG AAT TAG CTG TAT CGT CAA GGC ACT CT3’
25nc25nc 5'-bio- GGG CAG GAC CGG GCA GGA CC 3'
2.5
3.5
4.5
5.5
6.5
-400 -300 -200 -100 0
E/V vs. Ag/AgCl
I/µA
Elaboration du capteur
Glassy carbon electrode
Film Polypyrrole-ODN
1. Adsorption 2. Interactionnon-specific
3. Hybridation
Elaboration du capteur
Glassy carbon electrode
Film Polypyrrole-ODN
1. Adsorption 2. Interactionnon-specific
3. Hybridation
11 PyrPyr--ODn4500 ODn4500 PyrPyr
Selectivity of the responseSignal intensityReusable
Specificity of the response
ODN probes density
4.1.24.1.2-- Electrochemical biosensor for DNA hybridization detectionElectrochemical biosensor for DNA hybridization detection
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4.24.2-- Electrochemiluminescence for DNA hybridization detectionElectrochemiluminescence for DNA hybridization detection
hυ
e-
hυ
FluorescenceElectrochemiluminescence
MICAM twin potentialitiesImplementation and Detection
Time and spatial controlCan be quantified
Key point is theConductivity of the supportE
hυe-
Au
7 x 10 -2 cm 2 Avidin
Pyr-ODN/Pyr =1/20000 ABEI-biotin
Tampon borate pH=10, 50 µM H2O2
hυhυe-
AuAu
7 x 10 -2 cm 2 Avidin
Pyr-ODN/Pyr =1/20000 ABEI-biotin
Tampon borate pH=10, 50 µM H2O2
NN--(4(4--aminobutylaminobutyl))--NN--ethylIsoethylIso--luminolluminol
biotinbiotin
NHHN
S
O
NH
NH
NH
NH(CH2)4-CO-NH-(CH2)4 -N
O
O
O
O
C2H5
NN--(4(4--aminobutylaminobutyl))--NN--ethylIsoethylIso--luminolluminol
biotinbiotin
NHHN
S
O
NH
NH
NH
NH(CH2)4-CO-NH-(CH2)4 -N
O
O
O
O
C2H5
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ApiT8ApiT8
4.24.2-- Electrochemiluminescence on a chipElectrochemiluminescence on a chip
6
7
8
9 1
2
3
4
Pyr-ODN/Pyr = 1/4000
Pyrrole
0
500
1000
1500
2000
2500
0 2 4 6 8 10 12temps (s)
sign
al (
nb c
oups
/50m
s)
ECL plot 1
0
50
100
150
200
250
300
350
400
450
500
0 2 4 6 8 10 12 14
temps (s)
sig
na
l (n
bs
co
up
s/5
0m
s)
ECL plot 9
hυe-
Pyr-ODN/Pyr =1/4000
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5-Electrochemical detection
of DNA hybridization:The SECM approach
5-Electrochemical detection Electrochemical detection
of DNA hybridization:of DNA hybridization:The SECM approachThe SECM approach
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Sample or substrate
XYZActuator
xy
z
xy
z
Bipotentiostat
POT 2mA
POT 1nA
Reference
Counter electrode
Working 2-substrate
Working 1 - µelectrode
5.15.1-- SECM: working principlesSECM: working principles
MedR MedO
a
MedR MedO
a
MedR MedO dMedR MedOMedR MedO d
d
MedR MedOMedR MedO
Stationnary modeHemispheric diffusionShort time constantLateral resolution
i T > i T, ∞∞i T > i T, ∞∞
i T > i T, ∞∞i T > i T, ∞∞
i T,∞∞ = 4nFDCa
L=d/a
Positive feedback (F+)(recycling)
Negative feedback (F-) (diffusion hindrance)
GC mode(Collection of
generated species)
µelectrode
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5.25.2-- DNA hybridization modulate the SECM feedback modeDNA hybridization modulate the SECM feedback mode
Single strand spotSingle strand spot
Positive feedback modePositive feedback mode
Decrease of the apparent substrate reactivity toward Fe(CN)63-/4-
No change for in feedback current for Ru(NH)33+/2+
DNA hybridization detection due to reppelant electrostatic forcereppelant electrostatic force
3’ thiol modified ODN (20 mers) spotsAnionic redox mediator
W. Schuhmann et al, Angew. Chem. Intl. Ed43 (2004) 3482
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5.35.3-- DNA detection through guanosine oxydationDNA detection through guanosine oxydation
Xanthon CorpRaleigh-DurhamUSAH.H. Thorp et al,Anal Chem 72 (2000) 3764
SequenceSequence dependantdependant
Biochips imaging using SECMBiochips imaging using SECM
F. Zhou et al, J. Electroanal. Chem. 537 (2002) 95
a c
Ru2+ Ru3+ Ru2+ Ru3+
Ru2+ Ru2+
a c
Ru2+ Ru3+ Ru2+ Ru3+
Ru2+ Ru2+
(Inosine modified)
F+ F-
Positive feedback modePositive feedback mode
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Amplification through substrate recyclingAmplification through substrate recycling
InfineonGermany
5.45.4-- Enzyme labelling for localised hybridization detectionEnzyme labelling for localised hybridization detection
GOxGOx
e-
GC mode GOx labellingH2O2 localised detection
Glucose
O2 H2O2
Gluconolactone
Glucose
APPAPPAPPAPP
e-
p-APP
p-AP
p-APP
p-AP
C. Toth et al, Bioelectrochem.63 (2004) 207
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DRFMCDRFMCCEACEA
SynchrotronSynchrotronILLILL
Some pictures of Grenoble and CEASome pictures of Grenoble and CEA
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AknowlegmentsAknowlegments
B. Corso
G. Bidan
E. Fortin
A. Roget
T. Livache
M. Billon
H. Kim
L. GrosjeanC. BrakhaN. Lassalle
L. Blum C. Marquette
M.-L. Calvo-MuñozE. Dunot
S. Szunerits
A. Thaumoux
J.P. CorreiaL.M. Abrantes
J. Chane-TuneE.Vieil
J.P. Petit
M. DemeunynckL. Bouffier
B. Wang E. Descamps Y. Defontaine
G. Denuault, S. Evans