openmicrowell based biosensor with dielectrophorec … fine secondo... · openmicrowell based...
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OpenMicrowellBasedBiosensor
with
Dielectrophore4cCellManipula4on
andImpedanceMonitoring
Tutor:
Prof.RobertoGuerrieriPhDStudent:AndreaFaenza
2ndyearpresenta4on
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OutlineLab‐on‐a‐chipisanadvancedtechnologythatintegratesamicrofluidicsystemonamicroscalechip.
Itistheminiaturiza4onofalaboratorytoasmalldevice.The'laboratory'iscreatedbymeansofchannels,mixers,reservoirs,diffusionchambers,integratedelectrodes,pumps,valvesandmore.
Goals:• Automatestandardlaboratoryprocesses.
FastandcostefficientImproveddataquality
Increasedsepara4onability
• ImproveResearchinvariousareasoflifesciences(e.g.Immunotherapy,Stemcells,Cancerresearch,etc…)
Individuallycontrollingsinglecellsandpar4cles
Issues:
‐Incompa4bilitywithsmallvolumeandrare(1/106)cells(e.g.CellSorter)‐Cellrecoveryimpossibleortooslow(e.g.ClosedMicroCage)
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OurplaWormisabiosensorbasedonOpenMicrowelldesignedforHighThroughputrecoverywithintegratedelectrodesfor:
dielectrophore4ccellmanipula4onimpedancemonitoringwithsinglecellresolu4on
OurplaWorm:
Steps:
‐ Infusion:PhysiologicalSolu4on+cells(typicalflowrate:1ul/min)
‐ CellSor4ng
‐ ControlledCellDeliveryusingDEP
‐ Forcecell2cellinterac4onusingDEPfocusing
‐ Par4clescoun4ngusingimpedancemeasurements
‐ Cellrecovery
PlaWormdescrip4on Controlleddelivery Cell2cellinterac4onusingDEPfocusing Impedancemonitoring
Dielectrophoresis(DEP)
Thedielectrophore4cforcedependson:
• Themediumandpar4cles'electricalproper4es• Thepar4cles'shapeandsize• Thefrequencyandmagnitudeoftheelectricfield
Aphenomenoninwhichaforceisexertedonadielectricpar7clewhensubjectedtoanon‐uniformelectricfield.Thisforcedoesnotrequirethepar4cletobecharged.
Allpar4clesexhibitdielectrophore4cac4vityinthepresenceofelectricfields.
V+ V‐
ElectricField
‐‐‐‐‐+
++
‐
‐‐‐‐
‐
‐‐
‐‐
‐
‐
‐‐‐
+++
+
++++++
+++
pDEP
V+ V‐
ElectricField
‐‐‐
‐
‐++
+
‐
‐‐‐‐
‐
‐‐
‐‐
‐
‐
‐‐‐
++
+
+
++++++
+++
nDEP
εp > εm εp < εm
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Fluidicchanneldimensions(HxWxL) 50μmX400μmXDevicelength
Flowrate0.5‐1μL/min(LAMINARFLOW)
Par4clevelocity(cellwith10‐15μmdiameter) 100μm/sec(StokesDragForce)
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Mul4LayerKapton®FlexPCBdesign
Microfluidicinterfacefabrica4on Result:
Geometricalproper4es:Devicethickness200μmMicrowelldiameter70‐100μmMicrowellpitchStandardMicro4tercompa4bility(2.25mm,mul4pleandsubmul4ple)
PlaWormdescrip4on
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Microchannel
Microwell
Air
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Controlleddelivery
Focaliza4onStructures:
DEPForce+DragForce:
Biosensor
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Normallythefreefallofthepar4cleswithinthewellischaracterizedbya
randomposi4oningNOCONTACT! Bojom
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Cell‐cellinterac4onthroughDEPfocusing
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Animportantrequirementofthebiosensoristoforcethecell‐cellinterac4on
DEPFocusingThe par4cles are pushed awayfromtheboundariesof thewellwhere the electric field ismaximum and drop along thecentralver4calaxis
Electrode Amplitude(V) Frequency(HZ)
Top GND ‐
Middle GND ‐
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a)
b)
c)
d)
e)
f)
a)
b)
c)
d)
e)
f)
Par4clesdockatthesameposi4ononthemeniscusandarefoundincontactbetweeneachothers.FocusLevel
FocusLevel
FocusLevel
FocusLevel
FocusLevel
FocusLevel
Cell‐cellinterac4onthroughDEPfocusing‐Results
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Par4clescoun4ngusingimpedancemeasurementsModel:
BIOSENSOR
MUX32:1 MUX32:1 MUX32:1
8x4Padsexample
Labviewinterface
freqє[100KHz–1MHz]Amplitude=250mV
Sensingboard
1.I/VConversionandpreamp 2.Mixersanddivider
Systemschema4za4on:
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SensingBoardBlockDiagram
I/Vconversion PreAmplifier X
DividerLPF
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I/Vconversion PreAmplifier X LPF
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VoutαZmidZtop
VirtualGND
(I/Vstage)
SovwarePostProcessingFeaturesPreservingSmoothing
Filter
DetrendFilter
PeakDetector
Savitzky–Golaylocalpolynomial
regression
Mul4resolu4ondetrendfilter
Mul4parametercriteria
Electrode Amplitude(V) Frequency(HZ)
Top VirtualGND ‐
Middle VirtualGND ‐
Bojom 0.25 100KHz–1MHz
Worksindependentlyoftheaxisthecellsfollowduringthedescent
(Integra4onwithDEPFocusing)
Compensatesfornoisyphenomena,suchastemperaturedrivorconduc4vitychanges,typicallyfoundinopenstructuresatair‐liquidinterfaces
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Time (step= 0.1 s)
Time (step= 0.1 s)
Volta
gVe
(V)
Rel
ativ
e Va
riatio
n
WorstCasetest:Temperatureinduceddriv74messmaller
Rel
ativ
e Va
riatio
n
Time (step= 0.1 s)
Sta<s<cs*
OutputPeakVaria4on
0.84±0.57%
Pulsewidth 9.0±3.0s
*K562leukemiacells
Results:
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Conclusions:
Wepresented:OpenMicrowellBasedBiosensorforcell2cellinterac4onstudy,abletoworkatsinglecelllevelanddesigntoallowcellrecovery
ControlledcelldeliveryusingDEPforce
Cell2cellinterac4onforcedusingDEPfocusing
Systemmonitoringandcellcoun4ngusingimpedancemeasurements
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Publica7ons:
“Dielectrophore<ctrappinginmicrowellformanipula<onofsinglecellsandsmallaggregatesofpar<cles”M.Bocchi,M.Lombardini,A.Faenza,L.Rambelli,L.Giulianelli,N.Pecorari,R.Guerrieri.Biosensors(Vol.24,Issue5),January2009
“Biocompa<bleLab‐On‐SubstrateTechnologyPlaJorm”,T.Braun,L.Böjcher, J.Bauer,M.Bocchi,A.Faenza,R.Guerrieri,R.Gambari,K.‐F.Becker,E. Jung,A.Ostmann,M.Koch,R.Kahle,R.Aschenbrenner,H.Reichl,Proc.ECTC2009,SanDiego,CA,USA.
“Con<nuousimpedancemonitoringofsinglecellsdeliveredinopenmicrowellarrays”,A.Faenza,N.Pecorari,M.Bocchi,E.Franchi,R.Guerrieri,MicroTas2009,Jeju,Korea,November2009
Conferences:
Biosensor2008Shangai,China,14‐16May2008.
XNa<onalBiotechnologyCongressPerugia,Italy,17‐19September2008
ECTC2009‐ElectronicComponentsandTechnologyConferenceSanDiego,CaliforniaUSA,26‐29May2009.
MicroTas2009Jeju,Korea,1‐5November2009.
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