Description:

ThePL-chipwasspecificallydesignedforthequantitativeanalysisofplateletthrombusformationprocessinvolvingplateletadhesion&aggregation,granulesecretion,andthrombusgrowth.

Theplateletthrombusformationprocesscanbequantifiedandvisualizedbymeasuringflow-pressurewaveformandvideomicroscopy,respectively.

plateletthrombusformation

plateletthrombusformation(P2Y12antagoNIST)

Characteristics:

CharacteristicsofthePTF-assayusingPL-chip

• Single-usemicrochip(PL-chip)producedbyahigh-precisioninjectionmolding
• Smallamountofwholebloodsample(350μl）
• Simplesettingoftheoptionalwallshearrate(500-2500s-1)
• Dual-monitoringsystem(byflowpressurewaveformandreal-timevideoimaging)
• Easyoperationcontrolledbyacomputerinterlockingsystem

MeasurementPrinciple:

Abloodsampleisplacedinthereservoirthatisconnectedtoahigh-precisionmicropump.Themicropumpinfusesmineraloilintothereservoir,whichimpelsthesamplebloodthroughtheinletportandintotheflowpaththatleadstoacollagen-coatedanalyticalpathconsistingof25capillarychannels.

Theprocessofplateletthrombusformationintheflowchamberiscontinuouslymonitoredbyavideo-microscope(10x)positionedundertheflowchamberandbyapressuresensorthattrackspressurechangesintheflowpath.Theflowrateofthemicropumpcanbeeasilyadjustedasneededbasedonthedesiredexperimentalconditions(range:4-40μl/min).

Whenbloodflowsthroughtheanalyticalpathofthemicrochip,plateletsadhereandaggregateonthesurfaceofthecollagen-coatedcapillarychannels.Theplateletaggregatesgraduallyincreaseinsize,andintheprocess,occludethecapillary,resultinginanincreaseofflowpressure.

Parameters:

Flowpressureanalysis

WhenbloodflowsthroughtheanalyticalpathofthePL-chip,plateletsadhereandaggregateonthesurfaceofthecollagen-coatedcapillarychannels.Theplateletaggregatesgraduallyincreaseinsizeandeventuallyoccludethecapillary,resultinginanincreaseofflowpressure.Thus,flowpressurepatternsreflecttheplateletthrombusformationprocess.

Definitionsoftheparametersusedforquantificationoftheplateletthrombusformationprocess

Twospecificterms,T₁₀andAUC₁₀,areusedtoquantifyplateletthrombusformationinsidethePL-chip.

T₁₀(=timetoreach10kPa)isdefinedastheonsetofplateletthrombusformationandrepresentstheduration(sec)fortheflowpressuretoincreaseto10kPafrombaseline.

AUC10(=areaundertheflowpressurecurvefor10min)representstotalthrombogenicityandreflectsonsettime,aswellasgrowthandstABIlity,offormedthrombi.

Antiplateletagents

Ingeneral,directinhibitionofplateletadhesionandaggregationbyGPIbα-orGPIIbIIIa-inhibitorscausesashiftintheflowpressurewaveform(i.e.,delayofonset).Incontrast,inhibitionsofauto-andpara-crineactivationsmediatedbygranulesecretionbyaspirinandclopidogrelsuppressariseofthepressurecurve,andoftenshowparabolicflowpressurepatternsduetothereducedstabilityandsustainabilityofthrombi.

Thus,flow-pressurewaveforms,aswellasparametersofT₁₀andAUC₁₀,provideusefulinformationtoanalyzetheeffectsofantiplateletagentsandtodiagnosecongenitalandacquiredplateletdysfunctions.

AssayProtocol:

PlaceaPL-chipontheassaystageandletstandfor1-3mintoallowstabilizationofthetemperature.

Connectthereservoirtothenozzle,andPipette350μlofhirudin-treatedwholebloodintothereservoir.

Closethelid,andthenremoveitscap.

ConnectthereservoirtothePL-chip.

Pushthestartbutton.