AFM modes UniversityPressScholarshipOnline OxfordScholarshipOnline AtomicForceMicroscopy PeterEatonandPaulWest Printpublicationdate:2010 PrintISBN-13:9780199570454 PublishedtoOxfordScholarshipOnline:May2010 DOI:10.1093/acprof:oso/9780199570454.001.0001 AFMmodes PeterEaton PaulWest DOI:10.1093/acprof:oso/9780199570454.003.0003 AbstractandKeywords ThemanydifferentimagingmodesandexperimenttypesthatmodernAFMscancarry outexplainitspopularity.Theytransformahigh‐resolutionmicroscopeintoaversatile measurementstoolthatcandetermineaverywiderangeofsamplepropertieswith nanometreresolution.Thischapterdescribesthedifferencesbetweenthevarious imagingmodesavailable,suchascontact,non‐contact,andintermittent‐contactmodes. Thetheoryandpractices,aswellasthestrengthsandweaknessesofeachmodeare highlighted.Furthermore,non‐topographicalmodes,whichcanmeasuremechanical, (bio)chemical,magnetic(MFM),electrical(EFM)andthermalproperties,arediscussed. TechniquessuchasforcespectroscopyallowtheAFMtodirectlymeasuretheforceof interactionbetweensinglemolecules.OtherAFMtechniquescanevenbeusedtomodify samples,andthenimagetheresults.Examplesoftheuseofallmodesaregiven,tohelp Page 1 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes thereadertounderstandtheirpotential. Keywords:AFMmodes,topographica,lnon‐topographical,forcespectroscopy,contact,non‐contact, intermittentcontact,MFM,EFM TherangeofavailableAFMmodesandexperimentsareattheheartofmodernAFM.The widevarietyofexperimentsthatmaybeperformedwithanAFMmakeitaversatile, powerfultool.Initiallytheonlymodeavailableforimagingwascontactmode,andthis limitedthetypesofsamplesthatcouldbeexamined,thetypesofexperimentsthatcould beperformed,andthetypeofdatathatcouldbeproduced.Nowthereareaverylarge numberofpossiblemodesofoperationofAFM.Forexample,in1999,Friedbacheretal. attemptedtolistthenamesofalltheSPMmodesdescribed,andarrivedatmorethan50 terms[94].SomeoftheseSPMmodeswereSTMorSNOM(ScanningNearfieldOptical Microscopy),butthereareatleast20differentmodesofAFM.SNOMisanexampleof usingtheclosecontactandpositioncontroloftheAFMtomeasurepropertiesofthe surfaceotherthantopography(inthiscase,opticalproperties).Asshowninthischapter, manyofnewermodesinAFMarealongtheselines:techniquesthatusetheincredible resolutionprovidedbyscanningaprobeclosetothesurfacewithanAFMtomeasure differentpropertiesofthesamplesurfaceonthenanoscale.SNOM(alsosometimescalled NSOM,near‐fieldscanningopticalmicroscopy),isaverypowerfultechniquecombining near‐AFMresolutionwiththespectroscopicinformationthatisavailablebyusinglight‐ basedtechniques.However,SNOMisnotcoveredinthisbookbecausealthoughsome earlySNOMsinstrumentsweredevelopedbymodificationofAFMs,experimentsare nowgenerallycarriedoutwithspecializedSNOMinstruments,whichareratherdifferent fromanormalAFM.ForreviewsofSNOM,see[95,96].Atablecategorizingmajor techniquesinScanningProbeMicroscopyisshowninFigure3.1. ForthepurposesofmakingthisapracticalguidetoAFM,inthischapterweconcentrate ontechniqueslikelytobeaccessibleandofinteresttothereader.Thismeansthatwewill notdescribeindetailtechniqueswhicharenotattainablewithcommercialAFMswithout significantmodification,norcovermodesthathavebeendescribedbutnotwidely adopted.Somemoreadvancedtechniquesarecoveredintheapplicationssection, Chapter7. 3.1Topographicmodes ThebasisofAFMasamicroscopictechniqueisthatitmeasuresthetopographyofthe sample.AsdescribedinChapter1,thedatasetsgeneratedinthiswayarenot conventionalimages,asproducedbyopticalmicroscopy,butratheramapofheight measurements.Thesemaybelatertransformedintoamorenaturalisticimagewithlight shading,perspective,etc.tohelpuspicturetheshapeofthesample(thisprocessis coveredinChapter5).Inordertomaketheseheightmeasurements,avarietyofmodes havedeveloped,whichcanbedividedintothosemodeswhichmeasurethestatic deflection(p.50) Page 2 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes Fig.3.1. SummaryofthenamesofsomeSPM‐basedtechniques. oftheAFMcantilever,andthosethatmeasurethedynamicoscillationofthecantilever. Thedifferencesbetweenthemodesleadnotonlytodifferentexperimentalprocedures, buttodifferencesintheinformationavailable,differingsuitabilitiesforparticularsamples, andeventodifferencesintheinterpretationofthedata. 3.1.1Contactmode ContactmodeAFMwasthefirstmodedevelopedforAFM.Itisthesimplestmode conceptually,andwasthebasisforthedevelopmentofthelatermodes.Therefore understandingcontactmodehelpstounderstandhowtheothertechniqueswork. Althoughthelimitationsofcontactmodepromptedthedevelopmentofmodesthatcould examinedifferentsamplesindifferentenvironmentsandgivedifferentinformation, contactmodeisstillanextremelypowerfulandusefultechnique.Contactmodeiscapable ofobtainingveryhigh‐resolutionimages.Itisalsothefastestofallthetopographicmodes, asthedeflectionofthecantileverleadsdirectlytothetopographyofthesample,sono summingofoscillationmeasurementsisrequiredwhichcanslowimaging. InordertounderstandthewayAFMmodeswork,itisnecessarytouseso‐calledforce– distancecurves.Acartoonofasimpleforce–distancecurveisshowninFigure3.2.Asthe nameimplies,thesecurvesareaplotofforce(ontheyaxis)versusdistance(onthex axis).SuchacurveissimpletoacquirewiththeAFM.Itiscalculatedfromadeflection– distancecurvewhichiseasilymeasuredbymonitoringthedeflectionofthecantileveras thepiezoisusedtomovethetiptowardsthesample.Typically,atasetdeflectionlevel, thedirectionisreversed,andthetipwithdrawsfromthesample.Thisresultsina deflectionversusdistancecurve,whichmaybeconvertedtoaforce–distancecurve. Measurementofforce–distancecurvesisaverysensitiveandquantifiablewayto(p.51) Page 3 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes Fig.3.2. Simplifiedforce–distancecurveshowingcontact(repulsive region)scanningregime.Adeflection–distancecurve,whichisthe rawdatafromwhichaforce–distancecurveismeasured,hasa similarshape.Right:illustrationofprobebendingineachregime. determinetip–sampleinteractions,andisthebasisforseveralnon‐topographicAFM modes,suchasforcespectroscopyandnanoindentation.Moreinformationabouttheuse ofsuchcapabilitiesofforce–distancecurves,andhowtheyareconvertedfromdeflection toforceisgiveninSection3.2.1,andSection4.5,respectively. ConsideringtheapproachcurveshowninFigure3.2,whenthetipisfarfromthesample surface,thecantileverisconsideredtohavezerodeflection;asthetipapproachesthe surface,itnormallyfeelsfirstanattractiveforce,anda‘snap‐in’occurs,asthetip becomesunstableandjumpsintocontactwiththesurface.Astheinstrumentcontinues topushthecantilevertowardsthesurface,theinteractionmovesintothe‘repulsive’ regime,i.e.thetipisnowapplyingaforcetothesample,andthesampleappliesan oppositeforcetotip.Inthisregime,acombinationofcantileverbendingandsample compressionwillbeoccurringaccordingtotherelativecompliancesofthesamplesurface andAFMprobe.Ifthedirectionofmovementisreversed,theinteractionpassesagain intotheattractiveregime,andthetipstaysonthesurfaceuntilinstabilityoccursonce more,andthetipsnapsoffthesurface.Itiswithintherepulsiveregimethatcontact‐ modeimagingusuallyoccurs(forexample,atthepointlabelled‘set‐point’inFigure3.2). Inotherwords,incontact‐modeAFM,thetipoftheprobeisalwaystouchingthesample. Thishasthefollowingimportantimplicationsforcontact‐modeAFM: 1.Asaresultoftherepulsiveforcebetweenthetipandthesample,thesample maybedamagedorotherwisechangedbythescanningprocess. 2.Conversely,thetipcouldalsobedamagedorchangedbythescanning process. 3.Asthetipandsampleareconstantlyincontactwitheachotherasthetipmoves alongthesample,inadditiontothenormalforcetheyapplytoeachother,lateral forcesareexperiencedbybothprobeandsample. 4.Thecontactbetweenthetipandthesamplemeansthatthenatureofthe samplesurfacemayaffecttheresultsobtained.Thismeansthatthetechniquecan besensitivetothenatureofthesample. (p.52) Theforcesappliedtothesurfacebytheprobeincontactmodearegivenby Hooke'slaw: F = −k × D Page 4 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes F = −k × D (3.1) whereF=force(N),k=probeforceconstant(N/m)andD=deflectiondistance(m). Thebasisofcontact‐modeAFMisthatthemicroscopefeedbacksystemactstokeepthe cantileverdeflectionatacertainvaluedeterminedbytheinstrumentoperator.Thispoint isknownastheset‐point.Theset‐pointisoneoftheimportantcontrolparametersthatthe operatormustadjusttooptimizeimaging,andthereareequivalentparametersforall otherAFMimagingmodesaswell.Equation3.1showsthateitheraprobewithahigh forceconstant(onewithastiffcantilever),oragreaterdeflection(i.e.ahigherset‐point), willleadtoahigherappliedforce.BecausethefeedbacksystemoftheAFMcannothave instantaneousresponse,theverticaldeflectionwillactuallyvarysomewhatduringimaging (indicatedbytheredregionofthecurveinFigure3.2).Theamountitvarieswilldepend onthetopographyofthesample,flexibilityofthecantilever,scanningspeed,andhowwell thefeedbackcircuithasbeenoptimized.Optimizationoftheseparametersisdiscussedin Section4.2.TheAFMsoftwaremaydisplaythedeflectionsignalasalineplotasthetip passesoverthesample,orasanimage.Thedeflectionsignalincontact‐modeAFMisthe errorsignal,thatis,thesizeofthedeflectionisameasureofhowmuchthecantileveris deflectingbeforethedeflectionis‘corrected’bythefeedbackcircuitviaheight adjustmentbythepiezo.Therefore,intheidealsituation,therewouldbenocontrastin thedeflectionimage.Themorecontrastexistsinthedeflectionsignal,themore‘errors’ willbepresentintheheightimage,becauseregionsofhighcontrastinthedeflection imagecorrespondtoregionsintheheightimage,wherethefeedbackhasnotyet correctedforcantileverdeflection.However,usuallyitisnotpossibletohavethe feedbacksignalrespondperfectly,andthedeflectionsignalwillshowtheslopeofthe sample,becauseitisregionswherethereishighslope,ormoreprecisely,agreatrateof changeofslopewithdistance,thatgiverisetolargecantileverdeflections. Theimagingmodedescribedsofarisknownasconstant‐forcecontact‐modeAFM.Ifthe userturnsofffeedbackaltogetherwhileimaging,thentheyareeffectivelyusingconstant‐ heightcontact‐modeAFMratherthanconstant‐forcecontact‐modeAFM.Because constant‐forcemodeisbyfarthemostwidelyusedmode,ingeneralanyreferenceto contact‐modeAFMwillmeanconstant‐forcemodeunlessspecifiedotherwise,andthisis theconventionwefollowinthisbook.Inconstant‐heightmode,withnofeedbackactive, theimagesignalcomesentirelyfromcantileverdeflection,ratherthanfromthevoltage appliedtothezpiezo(whichwouldbetypicallysetataconstantvalue).Height measurementswillthereforerequirespecificcalibrationofthecantileverdeflection,to extractrealsampletopography.Constant‐heightmodeAFMdoeshavesomespecific applications:itcanbeusefulinconditionswherescanningiscarriedoutsofastthatthe feedbacksystemcannotcope[9,36].However,undertheseconditions,AFMisinfact actingratherlikeastylusprofilerasthetip–sampleforceisnotfullycontrolled.Typically, tocarryoutthesemeasurements,thefeedbacksystemisinitiallyusedtodeterminethe locationofthesamplesurface,andtheapproximatetopography,beforebeingturnedoff, orjustreducedtoaverylowlevel. Page 5 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes However,evenusingconstant‐forceAFM,thesoftwaretypicallywillallowtheuserto savethedeflectionimage,andsomeoperatorschoosetopublishthisimage,asitisoften a(p.53) Fig.3.3. Illustrationoftherelationbetweenheightanddeflection images.Lefttoright:height,right‐shadedheightanddeflection imagesofthesurfaceofamosquitoeye.Theheightimageshows howmuchthezscannermovestomaintaintheset‐point.The deflectionimageshowshowthecantileverbendsasitpassesover thesample,andisthesignalusedforfeedbackincontact‐mode AFM.Thisimageisverysimilartotheshadedheight. simplewaytoshowtheshapeofthesample,andmayevenshowfeaturesnotvisiblein theheightimage(whichcouldbeanindicationthatfeedbackwasnotoptimized). However,itisworthrememberingthatwherefeedbackwascorrectlyoptimized,the AFMheightimagewillalsocontainallthefeaturespresentintheerrorsignal.Onewayto showthisistoapplyashadingalgorithmtotheheightimage–thiseffectivelygivesthe derivativeoftheheightimage;theresultingimagewillbeverysimilartothedeflection image.AnexampleofthisisshowninFigure3.3.Notethatinthedeflectionimageshown inFigure3.3thez‐scaleisinvolts.Thez‐scalewasincludedhereforillustrative purposes,insuchimagesthez‐scaleisalmostcompletelymeaninglessscientifically–even ifconvertedtonanometres,thesizeofthedeflectioncouldeasilybechangedby adjustmentofthefeedbackparameters,andsoshouldalwaysberemovedfromthe imagebeforepresentation. Thedeflectionsignalisused,asdescribedpreviously,withthefeedbackparametersto determinehowthezpiezoelectricmustmovetomaintainaconstantcantileverdeflection (andhenceconstanttip–sampleforce).Theamountthezpiezomovestomaintainthe deflectionset‐pointistakentobethesampletopography;thissignal,plottedversus distance,formstheheightortopographyimageincontact‐modeAFM.Thereisathird signalwhichistypicallyavailableincontact‐modeAFM.Thisderivesfromthelateral twistingofthecantilever,andisthereforeusuallycalledlateraldeflection.Thissignalis typicallyusedduetoitsmaterialsensitivity,ratherthanasameasureofsample topography,anditisthereforecoveredinthenon‐topographicmodespartofthischapter (Section3.2.3.1).Theoriginoftheverticalandlateraldeflectionsignalsinatypicaloptical leverAFMset‐upisshowninFigure3.4.ThephotodetectorusedinopticalleverAFMs usuallycomprisesoffoursegments.Thedifferenceinsignalbetweenthetoptwoand bottomtwosegments,i.e.(A+B)–(C–D)givestheverticaldeflection(measuredinvolts, oramps),andthedifferencebetweentherightmosttwosegmentsandtheleftmosttwo segmentsgivesthelateraldeflection,i.e.(B+D)–(A+C). Itisworthnotingherethatincontact‐modeAFM,likemostoftheothermodes,two Page 6 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes versionsofeachdatatypecanbeavailable,thesebeingthedatacollectedintheleft‐to‐ rightdirection,andthereverseset,collectedintheright‐to‐leftdatadirection.By(p.54) Fig.3.4. Illustrationofhowthephotodetectordetectsverticaland horizontalbendingofthecantilever. conventionAFMimagesarepresentedwiththefastscanaxisdataappearinghorizontally, nomatterinwhichdirectiontheAFMtipwasscanned.Sotherewillbeforwardand backwardheight,verticaldeflectionandlateraldeflectionimagesavailable,meaningupto sixdatachannelsmightberecorded.Iftheinstrumentisequippedwithazaxis calibrationsensor,bothzvoltageandzsensorchannelsmightbeavailable,raisingthe totaltoeightchannels.Typically,whiletheprobescansovereachlineinbothdirections, onlyonedirectionwillbesaved.Thisisbecausetheheightdatainthetwodirections shouldbeidentical.Theverticaldeflectionimagesshouldbethesameonpartsofequal slope,andgiveoppositecontrastonregionsofchangingslope,buttheinformation availablefromthedatacollectedinthetwodirectionsiseffectivelythesame.Soformost channels,thereisrarelyanyneedtosavethedatacollectedinbothdirections,although itissometimesusefultoobservebothforwardandbackwardheightdatawhile optimizingscanningconditions,asdiscussedinSection4.2.Lateraldeflectiondatafrom bothdirectionsissometimessaved,tohelpunderstandfrictionalpropertiesofthe sample,whichisdiscussedinSection3.2.3.1. Applicability Contact‐modeAFMhasareallywiderangeofpotentialapplicationsandsomeoftheseare describedinChapter7.However,itispossibletosummarizesomegeneralcaseswhere contactmodeislikelytobechoseninpreferencetoothertechniques.Probablythebest reasontousecontactmodeisitshighresolution.Somedynamicmodescanalsoachieve extremelyhighresolution,butcomparedto,forexample,intermittent‐contactmode,the resolutionofcontactmodeispotentiallyextremelyhigh.Whatkeepsitfrombeingused morewidelyisthattheappliednormalforceleadstoahighlateralforceappliedtothe sampleaswell.Inthecaseofweaklyadsorbedsamples,orsoft,easilydeformedsamples, thiscanleadtoproblemsofsampledistortion,damage,orevenremovalfromthe substrate[97,98].Becauseofthis,ithasbeensuggestedthatcontact‐modeAFMisno goodforsoftsamples.Thisisnotthecase,astherearemanyreportsofsoftbiological samples,eveninahydratedstate,beingsuccessfullyimagedbycontact‐modeAFM(for examples,see[99–103]).Often,contactmodeischosentoimagesuchdelicatestructures whensub‐molecularresolutionisrequired[99,100,102,104].What(p.55) istrueis thatinambientconditions,acapillarylayerofwaterwillformbetweenthetipandthe surface.Oneeffectofthisisto‘pull’theAFMtipontothesurface,oftenapplyinganeven strongerforcethantheforceapplied(viatheset‐point)bytheoperator.Thusitiseasyto Page 7 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes unwittinglyapplyaverylargeforce(>nN)tothesampleincontact‐modeAFMinambient conditions.Inwater,theseforcesdonotexist,soitiseasiertoimagewithaverygentle force.Forthisreason,andduetosomecomplicationsofimagingindynamicmodesin liquids(seethenextsection),imaginginliquidisastrongpointofcontactmode.As mentionedpreviously,contactmodealsoworkswellinhigh‐speedAFM,andsomehigh‐ speedAFMset‐upsusethismodeexclusively[36]. 3.1.2Oscillatingmodes InthefirstpaperonAFM,BinnigandQuateacknowledgedthepotentialbenefitsof oscillatingthecantileverinanAFM,andcomparedtheresultsofusinganoscillating probewiththosefromcontactmode.Atthetimecontactmodegavefarbetterresults, probablyduetothenatureoftheprobeused[19].Althoughtheuseofoscillatingmodes wererevisitedshortlyafterwards[105],itwasseveralyearsbeforeoscillatingprobe modesbecamepopular,andforquiteawhilenearlyallAFMwascarriedoutincontact mode.TheprimarymotivationforusingoscillatingmodeinanAFMistotakeadvantageof thesignal‐to‐noisebenefitsassociatedwithmodulatedsignals.Thus,anAFMthathas oscillatingmodescanmeasureimageswithasmallprobe–sampleforce. Therearenowalargenumberofdynamicmodesofoperation,andevenmorenamesfor thosemodes.However,allofthesemodesarevariationsonatheme.Thecantileveris oscillated,usuallywithanadditionalpiezoelectricelement,andtypicallyatitsresonant frequency.Whentheoscillatingprobeapproachesthesamplesurface,theoscillation changesduetotheinteractionbetweentheprobeandtheforcefieldfromthesample. Theeffectisadampingofthecantileveroscillation,whichleadstoareductioninthe frequencyandamplitudeoftheoscillation.Theoscillationismonitoredbytheforce transducer(i.e.bytheopticalleverinmostAFMs),andthescanneradjuststhezheight viathefeedbacklooptomaintaintheprobeatafixeddistancefromthesample,justasin contact‐modeAFM.Theonlyrealdifferencesbetweenthevariousoscillatingmodes availableareinthesize(amplitude)oftheoscillationappliedtotheprobe,andthemethod usedtodetectthechangeinoscillation.ThegeneralprincipleofoscillatingAFMmodesis showninFigure3.5. Irrespectiveofthemanydifferenttermsusedtodescribethetechniques,thereare actuallyonlyafewkindsofconditionsusedinoscillatingimagingmodes.Theusercan decidetoseteitherasmalloralargeappliedoscillationamplitude,andsometimescan decidehowtodetectthechangeinprobeoscillation.Someinstrumentsmayonlyhave onedetectionschemeimplemented.Theinstrumentalset‐upschematicisshowninFigure 3.5.Anoscillatingsignalisgenerated,andappliedtothecantilevermechanically,suchthat theprobeisoscillatedclosetoitsresonantfrequency.Theoscillationoftheprobeis monitoredasitisbroughtclosetothesamplesurface.Thedetectedchangeinthe oscillation(whetherdetectedviaamplitude,phaseorfrequency),isusedinafeedback looptomaintaintheprobe–sampleinteractionconstant.Thechoiceofsmallorlarge amplitudehasaconsiderablepracticaleffect,asisillustratedinFigure3.6.Usingasmall oscillationamplitude(DenotedbythearrowA),itispossibletomaintain(p.56) Page 8 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes Fig.3.5. SchematicofgeneralizedoperationofoscillatingAFM modes,showinginstrumentalset‐up.Anoscillatinginputsignalis appliedtothecantilevertomaketheprobevibrateupanddown. Theactualmovementoftheprobewilldependonitsinteractionwith thesamplesurface.Theresultingoscillationinthecantilever deflectionismeasuredandcomparedtotheinputoscillationto determinetheforcesactingontheprobe. thecantileverintheattractiveregimeonly.Thistechniqueissometimesknownasnon‐ contactAFM,oralternatively,andperhapsmoreaccurately,asclose‐contactAFM(see Table3.1).Thistechniquehassomeadvantagesduetothelowprobetip–sampleforces involved,andisdiscussedbelowinSection3.1.2.1.Ontheotherhand,itcanbeseenthat ifalargeoscillationamplitudeisapplied,thentheprobewillmovefrombeingfarfromthe surfacewherethere'snotip–sampleinteraction,throughtheattractiveregime,intothe repulsiveregime,andback,ineachoscillationcycle(arrowB).Thistechniqueinvolves largeprobetip–sampleforces,socanbemoredestructive,butiseasiertoimplement. Thistechniqueiswhatwecallintermittentcontact‐modeAFM(andisalsoknownbymany othernames,someofwhicharegiveninTable3.1),andisdiscussedinSection3.1.2.2. Fig.3.6. DifferentoperatingregimesforoscillatingAFMmodes.A: withasmallamplitudeofoscillation,theprobecanbekeptinthe attractiveregime.B:withalargeroscillationtheprobemoves throughnon‐interacting,attractiveandrepulsiveregimes,resulting inintermittentcontact. (p.57) Table3.1.NomenclatureofsomeoscillatingprobeAFMmodes. Page 9 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes Detection Amplitude Low Amplitude Phase High Rarelyused IntermittentContactAFM(IC‐AFM), alsoknownasAC‐AFMorTapping Non‐contactAFM(NC‐AFM), alsoknownasclose‐contactAFM Rarelyused TypicallyanAFMdesignedforuseinairorliquidhaselectronicsthatcanmeasure changesinvibrationalamplitudeorphaseatapreselectedfrequency.Sotheinstrument operatorcanchoosetouseeitheroftheseforfeedback.Incombinationwithlargeor smallamplitudes,therearefourtypesofoscillatingexperimentavailabletomostAFM users,whichareshowninTable3.1. Itshouldbestressedthatthetwopossibleconditionsdescribedas‘rarelyused’inTable 3.1arenotunusable,justthattheyarenotcommonlyapplied.Phasedetectionisusually usedwithsmallamplitudes(close‐contactAFM),duetosomewhathighersensitivity,and amplitudedetectionisusuallyusedwithlargeamplitudes(intermittent‐contactAFM),but thesearenottheonlypossibleimagingmethods.Optimalimagingconditionsare sometimesdifficulttoestablish,anditmaybenecessarytotrydifferentamplitudesand detectionschemestofindtheidealconditions. Analternativetoamplitudeorphasedetectionisfrequency‐modulationdetection(FM‐ AFM),typicallyusedinultra‐highvacuumconditions(UHV‐AFM).FM‐AFMistypically appliedwithsmalloscillationamplitudesinthenon‐contactregime.TypicallyFM‐AFMis carriedoutwithaphase‐lockedloopdevice.ThistechniqueisunavailabletomostAFM usersduetotheneedforadditionalequipment,soitisnotcoveredindetailinthisbook. However,ithasbeendescribedindetail[106,107],andcomparedwiththeamplitude modulation(AM‐AFM)techniqueswediscusshereelsewhere[108]. 3.1.2.1Non‐contactmode/close‐contactmode OneofthegreatadvantagesofoscillatingmodesinAFMisthattheycandecreasethe sizeoftip–sampleforces,whilemaintaininghighsensitivitytothesampletopography.To achievenon‐contactAFM,thetipmustbecloseenoughtothesamplesurfacetoachieve thishighsensitivity,withoutpassingintotherepulsiveregimeusedforcontact‐mode AFM.Non‐contactAFMisthereforecarriedoutintheattractiveregime,asshownin Figure3.7. Byusingahighlystiffcantileverandmonitoringthedynamiceffectsoftheattractiveforce (i.e.thechangeintheoscillation)inthisregime,itispossibletomaintainthecantilever veryclosetothesurfacewithoutjumpingtotherepulsiveregime.Itispossibleto observechangesintheoscillationamplitudeandphaseinthisregime.Theseeffectsare causedbyachangeinthecantileverresonantfrequencywhichisinturncausedby forcesfromthesurface(normallyattractivevanderWaalsforces)actingonthetip.The Page 10 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes resonantfrequencyfarfromthesurface,ω0isgivenbyω0=c√kwherecisafunction ofthe(p.58) Fig.3.7. Operatingregimefornon‐contactAFM.Withasmall amplitudeandstiffcantilever,theprobecanoscillatewithinthe attractiveregimeonly. cantilevermass,andkisthespringconstant.Butanadditionalforceffromthesurface meansthatthenewresonantfrequencyω ′ O isgivenby: −−−−− w′0 = c√k − f ′ (3.2) wheref′isthederivativeoftheforcenormaltothesurface[109]. Theimportantpointhereisthateitherthechangeinamplitudeorthechangeinphase (whichactuallyderivesfromthechangeinfrequency)maybeusedinthefeedback circuittomaintainthetipatafixeddistancefromthesamplesurface.Thenamenon‐ contactAFMisactuallyquitemisleading.AllAFMmodesinvolvetheprobemovinginto theforcefieldofthesamplesurface,including‘non‐contact’AFM.Atthesortofdistances involved,itisimpossibletosayatwhichpointcontactoccurs.Furthermisunderstanding iscausedbythefactthatanumberofothernameshavebeenusedfordynamicAFM modes,andthereisnoclearconsensusonthecorrecttermstouse,sothereisgreat scopeforconfusion.Hereweusethetermnon‐contact‐modeAFMtomeanAFMcarried outintheattractiveregime,typicallyusingsmallamplitudesofoscillation.Section3.1.2.2 dealswithdynamicmodesthatpassintotherepulsiveregime,whichwechoosetocall intermittent‐contactmode. Non‐contact‐modeprinciplesofoperation Typically,non‐contactmodeiscarriedoutinamplitudemodulationmode,andtheerror signalmaybeeithertheamplitudeorphaseofoscillationofthetip.Toavoidthepossibility ofslippingintotherepulsiveregimewhichislikelytodamageorcontaminatethetip[110], ahigh‐frequencycantileveristypicallyusedwithω0intherangeof300–400kHz.In addition,smalloscillationamplitudesareused,oftenoftheorderof10nm[111].Aswith alldynamicmodesofoperation,scanningspeedisusuallylowerthanincontactmode, althoughthehighfrequenciesandsmallamplitudesmeanscanningspeedcanoftenbe greaterthaninIC‐AFM.WhenusedinUHVconditions,frequencymodulationisusually Page 11 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes used[108]. Applicability Non‐contact,orclose‐contactAFMisaverywidelyappliedtechnique,andcanbeused forimagingofalmostanysampleinAFM.Itiscurrentlyusedlessoftenthanintermittent (p.59) Fig.3.8. Possiblenon‐contactimagingconditionsunderambient conditions,withasamplecoveredinacontaminationlayer.Sucha layerexistsonmostsamplesinair.Inthefirstcaseontheleft,the probeoscillatesabovethecontaminationlayer.Inthesecondcase; right,theprobeoscillateswithinthecontaminationlayer. contactinambientconditions.However,withcare,itcanreplaceintermittentcontactin nearlyallapplications,andoftengivesbetter,andmoreconsistentresultsduetolower tipwear.Oneofthelimitingfactorsfornon‐contactmodeinairisthecontaminationlayer presentonmostsurfacesunderambientconditions.Ingeneral,thepresenceofthis layermeansthattheprobe–surfaceinteractionforcesaregovernedbythecapillary forcesbetweentheprobeandthecontaminationlayer.Fornon‐contactAFM,Theprobe maybevibratedintwodifferentdistinctregimesasitisscannedacrossthesurface,see Figure3.8.Inthefirstregime,theprobeisoscillatedabovethesurfaceofthe contaminationlayer.Thevibrationamplitudemustbeverysmallandaverystiffprobe mustbeused.Theimagesofthesurfacecontaminationlayeraretypically unrepresentativeofthesubstratetopographyandappeartohavelowresolution.Thisis becausethecontaminationfillsinthenanostructuresatthesurface.However,insome casesthistechniqueallowsthedeterminationofthelocationorshapeofliquiddropletson thesamples'surface,whichmaybedesirable[112,113].Inthesecondregimethe probeisscannedinsidethecontaminationlayer[110].Thistechnique,sometimescalled ‘nearcontact’,requiresgreatcaretoachieve.Again,thecantilevermustbestiffsothat thetipdoesnotjumptothesurfacefromthecapillaryforcescausedbythe contaminationlayer,andverysmallvibrationamplitudesmustbeused.However,high‐ resolutionimagesmaybemeasuredinthisregime.Non‐contactAFMfullyimmersedin liquidisalsopossible[114],anddelicatesamplessuchasDNAmoleculesorother biologicalsampleshavebeenimagedbyinthisway,andsuchmoleculesmaysufferless distortionwhenimagedlikethisthanwhenimagedbyintermittent‐contactmode[114– 116]. Usingultra‐highvacuum(UHV)conditions,FMdetectionhasadvantagesoveramplitude orphasedetection[117]andFMdetectioniswidelyusedforUHVnon‐contactAFM. Someamazingresultshavebeenshownforfrequency‐modulationbasednon‐contact AFMinultra‐highvacuum,includingtrueatomicresolution[118,119].Forinstance,the Moritagrouphaveshowntrueatomicresolutioninanumberofsystemswiththis Page 12 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes technique[106,118,120–122].Thesystemmustbeverystableforoperationtobe reliablewithouttheriskofjump‐to‐contact.Anexampleimageshowingtrueatomic resolutionbyNC‐AFMisshowninFigure3.9.Figure3.9alsoshowsarareexampleof usingNC‐AFMtoidentifyatomsonasurface.Forcespectroscopyisdescribedfurtherin Section3.2.1,withrespecttousingforcespectroscopyincontactmode.Butinthis example,unusualduetothemeasurementofforcecurvesinFM‐AFMmode,force spectroscopywasusedto(p.60) Fig.3.9. Examplenon‐contactAFMimages.Top:examplesofnon‐ contactAFMimagesinambientconditions(air)–individualDNA molecules(left)and1nmnanoparticles(right)[123].Bottomimages: non‐contactAFMinUHVconditionsforindividualatomidentification. Left:atomicallyresolvedNC‐AFMimageofSi,SnandPbatomson anSi(111)substrate–someatomsmaybedifferentiatedbasedon apparentsize,butidentificationisnotpossible.Middle:short‐range chemicalforcemeasuredovereachatomisdependentonthe chemicalnatureoftheatoms.Right:thesameimageasontheleft, withatomscolouredaccordingtothecolourschemeinthemiddle. Adaptedfrom[8],withpermission.(Acolourversionofthis illustrationcanbefoundintheplatesection.) identifytheattractiveforceaboveindividualatomswhichcouldbecorrelatedtotheir chemicalidentity[8].Furtherexamplesoftheapplicationsofnon‐contact‐AFMtoobtain atomicallyresolvedinformationaregiveninSection7.1.5. 3.1.2.2Intermittent‐contactmode AlthoughthefirstexperimentsindynamicAFMaimedtocarryoutnon‐contactAFM,it wasnotlongbeforetheadvantagesofusingadynamicmodethatallowstheprobeto touchthesample(thatis,passintotherepulsiveregime)werediscovered[97].For intermittent‐contactAFM,feedbackisusuallybasedonamplitudemodulation[108]and thetip–sampleinteractionpassesfromthe‘zero‐force’regime,throughtheattractive regime,andintotherepulsiveregime,asshowninFigure3.10. Thefactthatthetip–sampleinteractionmovesthroughallthreeregimeshasseveral importantimplications: Page 13 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes (i)Thereistip–samplerepulsiveinteraction,i.e.tipandsampletoucheachother, leadingtothepossibilityofsampleortipdamage,however: (p.61) Fig.3.10. Intermittent‐contactoperatingregime.Inthis mode,theAFMprobe'soscillationislargeenoughtomove fromtherepulsiveregime,throughtheattractiveregime, andcompletelyoutofcontactineachcycle. (ii)Duetothemovementofthetipperpendiculartothesurfaceasitscans,lateral forcesare(almost)eliminated. (iii)Thetippassesthroughthecontaminationlayer(seeFigure3.11). (iv)Tip–samplecontactalsoallowssomesensingofsampleproperties. (v)Thefeedbacksystemrequiresthecollectionofadequatedatatocharacterize thecantileveroscillationintermsofitsamplitude. Points(ii)and(iii)aboveexplainthepopularityofIC‐AFM.Thelateralforceswhichcan causegreatproblemsincontact‐modeAFMdonotaffectIC‐AFM.Ontheotherhand,the fundamentalinstabilityofnon‐contactAFMinair(duetooperationintheattractive regime,andthepresenceofthecapillarylayer)isovercome,makingIC‐AFMsomewhat simplertoachieve.InIC‐AFM,therestoringforceofthecantileverwithdrawsthetip fromthecontaminationlayerineachcycle,thusreducingtheeffectofcapillaryforceson theimage. Fig.3.11. Intermittent‐contact‐modeimagingconditionsinair.The probepassesthroughthecontaminationlayertotouchthe substratesurface,andoutagain. (p.62) Operatingprinciplesofintermittent‐contactAFM InIC‐AFMtheprobeisoscillatedwithalargeamplitude,typicallyintherangeof1–100 nm[108],andthefeedbackisusuallybasedontheamplitudesignal.Inmostcases,the probeisoscillatedbyanadditionalpiezoelectricelementattachedtotheprobeholder (seeChapter2),althoughitisalsopossibletoexcitethecantilevervibrationbyother methods,e.g.byanexternalmagnet,withamagneticallycoatedcantilever[124,125], Page 14 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes whichmayreducefluidvibrationwhenimaginginliquid.Infact,ratherthandrivingthe probedirectly,themostcommonexcitationmethodforfluidimagingistoexcitethe entirefluidcellholder,whichcausestheliquidtovibrate,acousticallydrivingthe cantilever[126,127].Often,inadditiontotheamplitudesignal,thedelayinthephaseof theprobeoscillationisrecorded.OscillationamplitudeandphaseareillustratedinFigure 3.12. Theamplitudeisreducedbythecontactwiththesamplesurface,andsoanamplitude set‐pointissetbytheuser,andtheamplitudeistheerrorsignalinIC‐AFM.Inasimilar waytodeflectionincontactmode,theamplitudesignalinintermittentcontactmaybe usedasanillustrationoftheshapeofthesample.Again,likethedeflectionsignal,the amplitudesignalshowswherethefeedbacksystemhasnotyetcompensatedforchanges insampleheight,soforbestheightdata,theamplitudesignalshouldbeminimized.An exampleimageshowingtherelationbetweenheightandamplitudedataisshownin Figure3.13.Notethatlikedeflectionimagesincontactmode,thezscaleofamplitude imagesinIC‐AFMisusuallyinvolts,unlessspecificallycalibrated.It'scommonpracticeto removethisscaleforpublicationasithasnopracticaluse. Inadditiontoheightandamplitudedata,thephase‐shiftmayalsobesavedasanimage. Thereasonwhysavingthisdataisusefulisnotobvious,andthisinformationwaslargely ignoredinearlyintermittent‐contactAFM.Infact,thephaseoftheoscillatingcantileveris stronglyaffectedbytheprobetip–sampleinteractions,soitcanbeausefulwayof distinguishingmaterials.AsaNon‐topographicmode,phaseimagingiscoveredinSection 3.2.3.2. Applicability Intermittent‐contactmodeisaverywidelyappliedtechnique,andiscurrentlythemost commonlyappliedtechniqueforimaginginair.Inliquid,IC‐AFMmodeisalsovery Fig.3.12. Illustrationoftheeffectofintermittentcontactonthe cantilevers'oscillation.Thefreeoscillation(solid)ismodifiedwhenin contactwithasurface(dashed)byareductioninamplitudeanda phaseshift. (p.63) Page 15 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes Fig.3.13. Intermittent‐contactAFMimagesofhumanredblood cells.Height(left)andamplitude(right)imagesshown. widelyapplied,althoughitissubjecttoanumberofdifficultiesspecifictooperationin liquid,namelythatmechanicalexcitationofthecantilevercanleadtoexcitationofthefluid andfluidcellaswell[128],andalackofclearunderstandingofthecontrastmechanisms [108,129,130].TheoperationofIC‐AFMmodeinliquid,aswellasinair,isdiscussedin Section4.3.Intermittent‐contactAFMisnotcommonlyappliedinvacuum,dueto restrictionsinbandwidthduetoincreaseofQinvacuum[117].Anextremelywiderange ofsampleshavebeenstudiedbyintermittentcontact‐modeAFM,someoftheseare illustratedinChapter7. Higherharmonicsimaging ArecentdevelopmentinIntermittent‐contactAFMistheuseofmodesofresonance otherthanthefundamentalone.Thismayeitherbebyapassivetechnique,bymeasuring thevibrationatthesehighermodes,caninvolveexcitationatmultiplefrequencies. AdditionofsuchcapabilitiestoanAFMisrelativelysimple,themainrequirementbeing thatalock‐inamplifiercapableofmonitoringtheveryhighfrequencies.Figure3.14shows illustrationsofthefirstfourmodesofabeam‐shapedcantilever.Therequirementfora high‐frequencyamplifierisbecausehighermodesofrealcantileversarelikelytohave extremelyhighfrequencies.Becausethemodesareanharmonic,thesecondmodeisnot necessarilyatdoublethefrequencyofthefundamental(i.e.f2≠2f1 ),butmaybeas highassixtimesthefundamentalfrequency[131].Inanycase,havingtwolock‐insis usefulbecauseitisadvantageoustobeabletomonitorbothf1andf2simultaneously. Fig.3.14. Illustrationsofthefirstfournormalresonancemodesofa beam‐shapedcantilever. (p.64) Thereasonforinterestinmonitoringthehighermodesofoscillationisthatithas beenshownthathighermodescanbemoresensitivetomaterialdifferences,particularly inthephasesignal[132].Garciaandco‐workershavestudiedthetheoryofthistypeof imaginginseveralworks[131,133,134]andexplainthatwhilethephaseshiftofthefirst fundamentalfrequencyissensitivetoenergyloss,thehigherharmonicscanbesensitive totip–sampleinteractionsthatconserveenergyaswell,explainingthecontrast improvementinhigherharmonicphaseimaging[134].Inrecentyears,morereports Page 16 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes haveemergedalsogivingfurtherexperimentalevidenceforthehighmaterialsensitivity ofthephaseshiftathighharmonics[135–137].Thishighsensitivityofthetechniquehas beenusedtoobtainhigh‐resolutionimagesinIC‐AFMevenofverysoftsamples[138, 139].ThesematerialsrequireverylowforceimaginginIC‐AFMmodetoavoiddamage, whichreducedthecontrastinthefundamentalmodetothepointwherenosub‐ moleculardetailswerevisible,butincreaseddetailswereavailableinthehigher oscillationmodes.Inaddition,ithasbeenreportedthatusinghigherharmonicsfor feedbackcanimproveimagingduetohigherQofthehighermodes[135]. 3.2Non‐topographicmodes EversincetheearlypapersonSTM,scanningprobemicroscopeshavebeenusedto obtainmorethanjusttopographicinformation.Inthoseearlyexperiments,thefirst reportsofascanning‐tunnellingspectroscopy(STS)experimentsweremade[140,141], whichconsistsoframpingthetunnellingvoltageandmonitoringthetunnellingcurrent withthetipheldfixedoveraparticularpartofthesamplesurface.Theuseoftheword ‘spectroscopy’hascontinuedintothefieldofAFM,where‘spectroscopic’techniquesare differentfrom‘microscopy’techniquesinthattheyprobepropertiesofthesampleother thantopography.Themostwell‐knownexampleisprobablyforcespectroscopy. 3.2.1Forcespectroscopy Forcespectroscopyinvolvesmaintainingthex‐ypositionoftheAFMprobefixed,while rampingitinthezaxis,tomeasurethedeflectionasthetipapproachesandretractsfrom thesamplesurface.Assuch,forcespectroscopyconsistsofsimplymeasuringforce– distancecurves,asshowninFigure3.15.Thegreatutilityofthistechniqueisthatthe AFMdirectlymeasurestheforcebetweenthecontactingatomsormoleculesontheend oftheprobeandsamplesurface,andasthecantilevermaybehighlyflexible,and deflectionsensitivitywithopticallever‐basedinstrumentsisveryhigh,single‐molecule interactionstudiesarepossible.Often,anAFMtipwillbemodifiedwithgraftedmolecules ofinterest[142–145],althoughsuchexperimentshavealsobeenreportedwithbare AFMtips[146,147],colloidalprobes[148–150](e.g.silicaspheres,whichmaybe themselveschemicallymodified),andevenmicro‐organisms[151,152].Thesurfaces probedhavebeenofevenwidervariety.Again,formolecule–moleculeinteractions studies,oftenaflatsubstratewillhavethemoleculesofinterestgraftedon[153],but alsocellmembranes[154],micro‐organisms[1155,156],wholelivingcells[157]anda widevarietyofsolidsurfacesincludingpolymers[158–160],metals[161],ceramics[162] andmorehavebeenprobed. Thereareanumberofexperimentalissueswhichmustbetakenaccountofinorderto performforcespectroscopy.Theseinclude:(p.65) Page 17 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes Fig.3.15. Amodelforce–distancecurve.AtpointA,theprobeisfar fromthesurface,atB‘snap‐in’occursasattractiveforcespullthe probeontothesurface.Theforcebecomesrepulsiveastheprobe continuestobedriventowardsthesample.Atsomeuser‐defined pointC,thedirectionoftravelreverses.AtpointD‘pull‐off’occurs astheforceappliedtothecantileverovercomestip–sample adhesion.Adhesiondataisusedforforcespectroscopywhileslope dataisusedfornanoindentation(Section3.2.2). (i)Thenumberofinteractingmolecules.Dependingonthetipradius,alarge numberofmoleculesarelikelytobeabletointeractwiththesurfaceatonetime. (ii)Orientationandaccessibilityofinteractingmolecules.Typically,theinvestigator wouldliketomakecomparisonsbetweenthemolecularinteractionsmeasuredat thesurface,andresultsfromsolutionstudies,butthegraftingofmoleculestothe tipmayaffecttheresults. (iii)Thespeedofapproachandwithdrawalofthetipforthesurfacewillaffectthe results. (iv)Experimentalenvironment.OneadvantageofAFMisthatitmaybecarried outinalmostanyenvironment.Formostchemicalandbiologicalworkitisuseful tocarryouttheexperimentsinliquid.Itissimplethentochangethecomposition oftheliquidtoseehowitaffectstheresults.Forexample,toprove antibody/antigeninteractions,commonlyblockingantibodiesareinjectedinto solution,afterwhichforcesmaydisappeartozero[163]. (v)Statisticalvariationinresultsistypicallyverylarge.Thismeansincreased experimentaltime,whichisnotnormallyaproblem,aseachforcecurvetypically takeslessthan1secondtoacquire,butinadditionaverylargedatasetis typicallygenerated,andalotofdataanalysisislikelytoberequired. Inreality,theresultsfromforcespectroscopybetweenmoleculesrarelylookmuchlike thecartooninFigure3.15.Usually,specificforcesbetweenmoleculesleadtomuchmore complicatedresults.AnexampleisshowninFigure3.16.Intheblue(retract)curve, severaltypicalfeaturescanbeseen.Oneisthealmost‐flatregionlabelleda.Inthisregion, polymerchainslinkingthemoleculestotheAFMtipwereunfolding.During(p.66) Page 18 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes Fig.3.16. Anexampleofrealforcespectroscopydata:curves measuredonM.xanthuscells.Theredtraceistheapproach,and theblueistheretractcurve.Reproducedwithpermissionfrom [164].Copyright2005NationalAcademyofSciences,USA. unfolding,onlyveryweakbondsarebroken,sothereareonlysmallverticaldeviationsin thetrace.Atb,theprobeappliedsufficientforcetobreakthebonds,asthemolecule breaksawayfromthereceptor.Notethatatthispoint,asingleverticalmovementmaybe expected,butthestepisstaggered,indicatingthatmultiplebondsarebroken,andonly atpointcisthetipfinallyfreeofmoleculeslinkingittothecellsurface.Inacasesuchas this,itisnecessarytodecideiftheverticaldistance(i.e.theforceofadhesion),seenat pointb,representstheadhesionofonemolecule,thatoftwomolecules,orofan unknownnumber.Thisiswhyitisdifficulttoautomatedataanalysisinforce spectroscopy,andthiscombinedwiththetypicalrequirementtocollecthundredsofdata points,meansdataprocessingforsuchexperimentscanbeverytime‐consuming.Some waystoimprovethesituationincludereducingthechanceofmultipleinteractionsinthe firstplacebyforexamplespacingthegraftedmoleculesoutonthetip,orlookingfor multiplesofsingleforcesinthe‘spectrum’offorcesmeasured[144]. Itcanbeusefultoperformforcespectroscopyinagrid‐likepatternoverthesample, leadingtothepossibilitytolocatespecificchemicalgroupsonasamplesurface[146,160, 165].Itisimportant,however,torememberthatevenhighlyspecificmeasurementslike adhesion–forceinteractions,maybeaffectedbysampletopography[159].Inthismode, forcespectroscopyissometimestermedchemicalforcemicroscopy[166].Amajor applicationofforcespectroscopyisproteinunfolding,whichusestheAFMforce sensitivitytoprobemechanicalunfoldingoflargeproteinmolecules,abiologically importantprocess,whichiscoveredinSection7.3.5.1. 3.2.2Nanoindentation IfinsteadofmeasuringthedataastheAFMwithdrawsfromthesamplesurface,we recordthedatameasuredasthetipcontactswithandpressesontothesamplesurface, wearecarryingoutadifferentexperiment,callednanoindentation.Anothertechnique knownasnanoindentationexists[167],whichusesadedicatedmachinetomeasureload– displacementcurvesasahardindenter(forexamplediamond)pressesintoasample. Typically,suchinstrumentsaredesignedtocreateaseriesofindents(holes)inasample, andallowthemeasurementofthesizesoftheindents(by,e.g.lightmicroscopy),andare sensitivetoforcesinthemicronewtonrange.Bycarryingoutan(p.67) analogous Page 19 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes experimentusingAFMwehavesomeadvantagesandsomedisadvantages.Theseare summarizedbelow. AdvantagesofAFM‐basednanoindentation •Highloadsensitivity–loadsensitivitymaybeaslowaspiconewton, althoughevenforsoftmaterialstherequiredsensitivityisnotlikelytobe greaterthanananonewton. •Inbuiltabilitytomeasuretheindentscreated,athighresolutioninx,yand z(seeFigure3.17). •Highpositioningresolution–i.e.wecanchoosesmallregionsofasample,or performtheexperimentonverysmallsamples. DisadvantagesofAFM‐basednanoindentation •Non‐perpendicularprobeapproach–quantitativenanoindentationrequires theindentertoapproachthesampleperpendicularly,whichisnotthecase normallyforAFM.Thisproblemcanbeovercome,withcare. •Non‐linearzpositioning.Unlessthesystemisequippedwithlinearizationin thez‐axisthiscancausesomeseriousproblems. •Thesystemmustbecalibratedtoextractrealforces. Fornanoindentationonhardmaterialsitisnecessarytouseaverystiffcantileveranda hardprobe.Typically,onemightuseacantilevermachinedfromsteel,withadiamondtip gluedtotheend[168].Suchleversmaybeappropriatetoperformnanoindentationand canbecapableofimagingthesample,buttypicallygiverelativelylow‐resolutionimages; ontheotherhand,theyareabsolutelynecessarytoindenthardmaterialsuchasmetals. ManyauthorshavealsocarriedoutnanoindentationwithnormalAFMprobes[168–172], butitisnecessarytocharacterizethetipradiusandcantilevercarefullyforquantitative results.Oneadvantageofsuchanapproachistheabilitytoselectfromawiderangeof springconstants;thehighlystiffnanoindentationcantileverspreviouslyreferredtoare inappropriateforsoftsamples.Onecommonapproachtosimplifytheproblemoftip radiusdetermination(seeChapter2)fornanoindentationmeasurementsistousea colloidalprobe,i.e.touseanormalAFMcantileverwithoutatip,butwithasmallspherical particleinitsplace[150,173].Ifnanoindentationexperimentsarecarriedoutinagrid patternoverthesamplesurface,thenit'spossibletodeterminethespatialvariationof hardnessandsoftness[158,174,175].Dataanalysisfornanoindentationisoftenmade bymodellingtheindentationviatheHertzmodel,whichrequiresknowledgeoftheshape ofthetip,andassumesonlyelasticcompressionsofthesampletakeplace[162,176].For morediscussionofdatatreatmentfornanoindentationseereferences[168,176,177]. Applicability DespitethequantificationissuesassociatedwithcarryingoutnanoindentationusingAFM, ithasbeenwidelyapplied.Itisparticularlyusefultolookatrelativehardnessand softness.Forexample,itcangiveanideaaboutdifferencesinhardnessandsoftnessin Page 20 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes differentpartsofasampleWithnanoindentationmapping,themeasurementscanbe madequantitative,whereasformanyothertechniquessuchasphaseimaging(see Section3.2.3.2),itishardtoknowifdifferencesareduetomechanicaloradhesive propertiesofthesample.Thereforenanoindentationhasbeencommonlyusedtostudy heterogeneousmaterialssuchaspolymercomposites[158,181,182].Furthermore,the highpositioningaccuracymeansit'spossibletolookatsmallfeaturesnotpossibleby traditionalnanoindentation,(p.68) Fig.3.17. Examplesofnanoindentationmeasurementswiththe AFM.Left:force–distancecurvesmeasuredwiththeAFMon individualbacteria.Blackcurves:typicaldatameasuredon untreatedandtreatedBacillusvegetativebacterialcells.Red curves:datameasuredonBacillusspores.Thedatashowedthat thetreatmentmadethecellssofter,butthesporesweremuch harderthanthevegetativecells[178].Right:AFMimageofan indentationmadebyadedicatednanoindenter.Theindentationisin amagnesiumoxidecrystal,andtheimageshowstheindentation (blacktriangle)pile‐up–materialpushedoutofhole(whitefeatures attrianglecorners),andalsoshowslong‐rangedislocationsinthe crystalstructure(diagonaldiscontinuities)[179].Reproducedwith permissionfrom[180]andkindpermissionfromDrC.Tromas. forexampleindividualmicro‐organisms[169,183](seeFigure3.17),livingcells[176, 184]ormicro/nanoparticles[185–187].Somemoreexamplesofapplicationsof nanoindentationaregiveninChapter7. 3.2.3Mechanicalpropertyimaging Nanoindentationisaveryusefultechniqueformechanicalcharacterizationbecauseofthe possibilitytocollecttrulyquantitativedataonthemechanicalresistanceofsamples. Howeverithasseveraldrawbacks,includingthecomplicateddataanalysis,andits relativelyslowdataacquisition.Theverylowrateofdataacquisitioncomparedtonormal imagingAFMmodesisamajordrawback.Foranimagewith512×512datapoints,afull setofnanoindentationdatawouldrequiremanyhourstocollect,leadingtoproblemswith thermaldriftofthesample.Forthisreason‘imaging’typestudieswithnanoindentation tendtobeusedonlyatverylowresolutions(100×100datapointsorless).Onewayto overcomethislimitationistomeasuretheinteractionoftheprobewiththesample surfacewhileitacquirestopographicaldata,andusethisinformationtoderivemechanical informationaboutthesamplesurface.Thishastwoadvantages,firstly,dataisacquiredat amuchfasterrate,andsecondly,themechanicalinformationcollectedmaybecorrelated directlywiththemeasuretopography.Thereareanumberofmodeswhichacquire mechanicalinformationaboutthesamplesurfaceinthisway,andtheyaredescribedin thefollowingsections. Page 21 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes 3.2.3.1Lateralforcemicroscopy AsdescribedinSection3.1.1,incontactmode,theverticaldeflectionofthecantilever, measuredasthedifferenceinsignalbetweenthetopandthebottomofthesplit photodiode,(p.69) isusedasthefeedbacksignal.However,ifwecomparetheleft‐and right‐handsidesofthesplitphotodetector,weobtainthelateraldeflectionsignal.When measuringthissignal,thetechniqueissometimescalledlateralforcemicroscopy,or LFM.Thereasonwhymeasuringthiscanbeusefulisthatthissignalcontainsinformation aboutthemechanicalinteractionoftheprobetipwiththesamplesurface.Thelateral twistingofthecantileverisameasureofthefrictionencounteredbythetipasitscans overthesample.Thus,thissignalissensitivetothenature(shapeandfrictional properties)ofthesurface.Forthisreason,LFMissometimesalsocalledfrictionforce microscopy(FFM),andthelateralsignalissometimesreferredtoasthefrictionsignal, althoughthesignalobtainedlaterallycontainsmoreinformationthanjustthefrictionfelt bythetip.Itisimportanttobearinmindthatthelateralbendingiscoupledwithvertical bendingofthetip,andcontainsinformationabouttheshapeofthesample,aswellasits material,becausefrictiondependsontheslopethetipistravellingalong[77,188]. However,usingthistechniqueitispossibletogetquantitativeinformationaboutvariation insampleproperties.SomeexamplesofthisareshowninSection7.1.4.Adiscussionon calibrationoflateralsignalsisincludedinSection4.2. Asmentionedpreviously,itisnotnormallynecessarytomeasureAFMheightsignalsin morethanonefastscanningdirection.Thesituationinthecaseofthelateraldeflection dataissomewhatdifferent.Thelateraldeflectionsignalwillnormallyalwaysbedifferentin thetwodirections,asthecantileverwilltwistbyacertainamountassumingthereissome measurablelateralcomponenttothetip–sampleforce(i.e.friction).Therefore,evenon perfectlyflat,homogeneoussamples,thetwoimageswillbedifferentfromeachotherin themagnitudeandpossiblysignofthesignal.Ingeneral,changesofslopewillaffect forwardsandbackwardsscansoppositely,andchangesinfrictionduetomaterial contrastwillgivegreaterorsmallerdifferencebetweentheforwardandreversescans. ThisisshownschematicallyinFigure3.18. FromFigure3.18itispossibletoseethatchangesinslopeandchangesinmaterial contrasthavedifferenteffectsuponthelateraldeflectionsignal.Iftheusersubtractsthe left‐to‐rightandright‐to‐leftsignalsfromeachother,inthecaseoftheslopechange,the resultwillbeasignalwithalmostnocontrast.However,inthecaseofthematerialfriction change,theresultingsignalwillbesensitivetothesamplefriction.Largerfrictionwillgive agreaterdifferencebetweentheforwardandreversescans,whilelowerfrictionwillgive asmallerdifference.Thus,collectingbothforwardandreversedirectionscansand subtractingtheminLFMcangiveusefulinformation[160,189]. 3.2.3.2Phaseimaging ‘Phaseimaging’inAFMreferstorecordingthephaseshiftsignalinintermittent‐contact AFM.In1995forthefirsttime,thephasesignalwasdescribedasbeingsensitiveto variationsincomposition,adhesion,friction,viscoelasticityaswellasotherfactors[190]. Page 22 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes Thenin1996GarciaandTamayosuggestedthatthephasesignalinsoftmaterialsis sensitivetoviscoelasticpropertiesandadhesionforces,withlittleparticipationbyelastic properties[191].Ithasbeenacommonassumptioneversincethatphasecontrastwill showadhesionorviscoelasticproperties[192,193].Infact,asshownintheexamplesof phasecontrastinFigure3.19,phasecontrastfrommaterialpropertiesisseeninawide varietyofsamples,butalsoreflectstopometricdifferences(differencesinslope).Thisis becausethephaseisreallyameasureoftheenergydissipationinvolvedinthecontact (p.70) Fig.3.18. Schematicoflateralforcesignalsrecordedonasample withvariationsintopographyonly(top)andinmaterialfrictiononly (bottom).Darkercoloursrepresentmaterialwithhigherfriction. Notethatinthecaseoftopographychanges(upper),thedifference betweentheforwardandbacklateraldeflectionsignalsisconstant; formaterialcontrast(lower),thedifferencechanges. betweenthetipandthesample[194–196],whichdependsonanumberoffactors, includingsuchfeaturesasviscoelasticity,adhesionandalsocontactarea[197].Ascontact areaisdependentontheslopeofthesample,thephaseimagealsocontainstopographic contributions,sounambiguousinterpretationofcontrastinphaseimagesisbestleftto flatsamples.Eveninsuchcases,understandingofthecontributionoftheindividual factorstothephaseshiftisnottrivial.Formoredetailsonthistopic,thereaderis recommendedtoreadtheexcellentandcomprehensivereviewsbyGarcia[108,197]. Despitethecomplicationsinvolvedininterpretation,phasecontrastisoneofthemost commonlyusedtechniquesfor‘mechanical’characterizationofsamplesurfaces,probably duetothe(p.71) Page 23 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes Fig.3.19. ExamplesofphasecontrastinIC‐AFMondifferent samples.Top:atriblockcopolymertopography(left)barelyshows heightdifferencesforthedifferentphases.Thephaseimage(right) showsclearcontrast.Bottom:Langmuir–Blodgettfilmonmica,the hightopographyregion(themonolayer)hasahigherphasecontrast thanthemicainthephaseimage.Thisimageshowshowtheedgesof thesephasesalsoshowdifferentcontrastinthephaseimage,dueto changesintip–samplecontactarea. popularityofIC‐AFM,andthefactthatobtainingthedataisverysimpleanddoesnot requirepost‐processingofthedata. 3.2.3.3Otherdynamicmodes Anumberoflesscommonlyusedoscillatingmodeshavebeenreported[198,199],these aretypicallyvariationsonIC‐AFM,designedtomakesimultaneousacquisitionofsample propertiesandtopographysimplerormorequantitative.Anexampleofthisisjumping modeAFM[198,200–204].ThisisavariantofIC‐AFM,thedifferencebeingthatin jumpingmode,themovementalongthefastscanaxisisdiscrete,ratherthancontinuous, andtheelectronicsaresetuptorecordthecantileverdeflectionatspecificpointsalong theforce–distancecurveduringeachoscillation.Theadvantageofsuchatechniqueis thatif,forinstance,thepointsrecordedareequivalenttopointsaandbinFigure3.2,the tip–sampleadhesionmaybeobtained,orslopedata(seeFigure3.15)couldberecorded toqualitativesamplestiffness.Theadvantageofthisparticularmodeisthattherelatively high‐speedscanningofIC‐AFMcanbecombinedwiththeacquisitionofsuchdata.Thisis alsotheaimofpulsed‐forcemode[199,205–207],whichoperatesinaverysimilarwayto jumpingmode,althoughfastscanaxismovementiscontinuous,likenormalIC‐AFM.As (p.72) Page 24 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes Fig.3.20. Exampleofpulsedforcemode.Thesampleisa polystyrene‐polymethylmethacrylateblend.A:topography,B: adhesion,bothmeasuredsimultaneously.Notethebrightborders betweenthephasesareduetoincreasedtip–samplecontactarea, andtheadhesionimageisinagreementwiththatmeasuredbyforce spectroscopy[159].Reproducedfrom[206]withpermission. withjumpingmodeAFM,amajoraimofpulsed‐forceAFMistoobtainadhesiondata [208],butcollectionofotherdatapointscanagainleadtosamplestiffnessdata[199].An exampleoftheresultsfrompulsedforcemodeisshowninFigure3.20. 3.2.4Magneticforcemicroscopy ThepotentialofusingAFMtomeasuremagneticpropertieswasrealizedquiteearlyin thehistoryofAFM[105,209,210].Magneticfieldsdecayquicklywithdistance,soin ordertomeasurelocalpropertiestheprobemustbeveryclosetothesurface,hence theapplicabilityofAFM.Themosttypicalexperimentcarriedoutisknownasmagnetic forcemicroscopy(MFM)[211].Inthismode,thepresenceanddistributionofmagnetic fieldsismeasureddirectly,byusingamagneticprobe.Typically,theseconsistof standardsiliconcantileverswithathinmagneticcoating.Typicalmaterialsusedforthe coatingincludecobalt,cobalt‐nickelandcobalt‐chromium[212].Theadditionofsuch coatingscanhavetwodetrimentaleffectsonthecantilever:firstlythesematerialsare typicallysofterthantheunderlyingsilicon,andthusmayincreasewearrate,and secondly,anycoatingaddedtotheendofthetipwillincreasetheradius,andthus decreasetheresolutionoftheexperiment.Typically,magneticforcesareordersof magnitudelowerthanothertip–sampleforceswhenincontact,andthusitisusefulto measurethemwiththetipatacertaindistance(oftheorderof5–50nm)fromthe surface,thusreducingtheinterferencefromshort‐rangeforces.Thiscanbecarriedout inanumberofways[213],someofwhichareillustratedinFigure3.21.Thesetechniques allhavesomepracticaladvantagesanddisadvantages,butarebasicallyvariationsona theme.In‘lifting’‐typemodes,thetopographyofthesampleismeasuredfirst,followedby raisingtheprobe,andscanningagaintocollectthemagneticdata.Onemethodisto collectanormaltopographyscan,andthenchangethezset‐pointtolifttheprobefrom thesurfaceandcollecta‘magneticimage’(p.73) Page 25 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes Fig.3.21. SchematicsofvariousimplementationsofMFM.A:lifting probebetweentopographyandMFMimages.B:Bardmethodof liftingleverbetweenscanlines.C:zset‐pointoscillation.D:Hosaka methodofmovingprobeclosetosurface,andrecordingMFM signalatvariouspointsforeachheight. (Figure3.21A).Thisworkswellforflatsamples,butispronetoproblemsoffeaturesfrom thesampletopographyappearingintheMFMimage,andalsotoproblemsfromthermal drift.AsdescribedbyBard[214],animprovedmethodistorecordthesample topographyfirst,thenlifttheprobe,andmeasurethelong‐rangeforceswhilefollowing theshapeofthetopography,butatacertain‘liftheight’.ThisisapplicabletoSTM,EFM (seethefollowingsection),orMFM.Typically,thisiscarriedoutinalternatescanlines, allowingthetopographydatatobeincludedinthesecond,magneticscanline,meaning theprobecanstayapproximatelythesamedistanceabovethesample,evenwithchanges intopography(Figure3.21B)[215].It'salsopossibletochangethezset‐pointwhile scanning,meaningtheprobewillbeconstantlymovingtowardsthesampletocheckthe topography,andthenawayagaintoregistermagneticfieldinformation(Figure3.21C). Finally,inthemethoddescribedbyHosaka[216],ateachpixeltheprobeisliftedabove thesurface,andthefieldismeasuredatseveralpointsastheprobeisloweredagain (Figure3.21D),toobtainamagneticfieldgradient.Theprobeisthenmovedtothenext lateralpoint,liftedagain,andsoon.Thismethodisprobablytheleastpronetothermal drift,butisratherslowtoimplement.Whichevermethodisused,liftingthetipfromthe surfacereducesresolution,andresolutioninMFMistypicallynogreaterthan30nm laterally[212]. Fortheseliftingmodestowork,ithelpsifthereislittlesampledrift,ortohavelinearized scanners.Typically,MFMiscarriedoutinoneofthedynamicmodes,andthemagnetic effectsonthecantileveraredetectedviaphaseshift,buttheymayalsoaffectthe oscillationamplitude.Unfortunately,evenatliftheightsofseveraltensofnanometres fromthesamplesurface,shortrangeforcesotherthanmagneticinteractionmayaffect thecantileveroscillation,givingafalseindicationofmagneticcontrast[217],aneffect which(p.74) issometimesoverlooked.Onewaytoovercomethisproblemistocarry outtwoscanswiththecantilevermagnetizationorientationinoppositedirections,and subtractthemfromeachother.Non‐magneticforcesshouldthencancelout,leaving typicallyasigmoidally‐shapedcontrastinthelinesscanswheremagneticinteractiontook place[218].AnexampleimageobtainedinMFMviatheBardmethodisshowninFigure 3.22. Page 26 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes AlthoughMFMisarelativelysimpletechniquetoobtainmagneticcontrastatahigh resolution,quantificationofMFMsignalsiscomplicated,andwhentryingtomeasurethe magneticdomainsonasoftmagneticmaterial,thedomainsontheprobecancausea changeinthedomainstructureonthesurface.Readersinterestedinmoredetailonthe issuesinquantificationofMFMsignalsaredirectedtotheworkofProkschetal.[218, 219].Itisworthpointingoutherethatthereareavarietyofothermagnetic characterizationtechniquesusingtheAFM,suchasMRFMthatinvolveconsiderably moreequipmentthanacommercialAFM[220],soareoutsideofthescopeofthisbook. Applicability TheinitialinterestinthestandardMFMtechniquegrewlargelybecauseofthepotential industrialapplications.Thedatastorageindustryislargelybasedaroundcreationof magneticnanodomainsofthesizerangeofafewhundredsofnanometres,andthereis noothertechniquetoaccuratelymeasuresuchfeatures.ThereforeMFMhasseenmuch useindustrially,particularlyindatastorageapplications[210,213].Morerecently, magneticnanoparticleshavebecomethefocusofintenseinterest,andtheseareanother fieldwhereMFMcanbeofgreatuse[221].Theverysmallmagneticmomentofthe smallestparticlescanpresentachallenge,andmuchworkhasbeencarriedouton particlesofca.50–100nm[221]butitshouldalsobepossibletoexaminethemagnetic fieldfromparticlesassmallas20nm.SomemoredetailsofindustrialapplicationsofMFM aredescribedinChapter7. Fig.3.22. ExampleMFMimages.Left:topographyofmagnetictape sample.Right:MFMimageofthesameregion,showingmagnetic fieldsaboverecordeddatabitsonthetape.Bothare10μm×10 μmimages. (p.75) 3.2.5ElectricforcemicroscopyandscanningKelvinprobemicroscopy Electricforcemicroscopy(EFM)referstoatechniqueanalogoustoMFMwhichenables themeasurementofelectricalfieldswiththeAFM,ratherthanmagneticfields. Essentially,thetechniquecanbeappliedbycarryingoutexperimentsinaliftingmodeas describedabove,butwithoutamagneticcoatingonthecantilever.Astandardsiliconor siliconnitridecantilevermaybeusedforsimpleEFMimaging,althoughconductive (metal‐coated)tipsarerequiredforread/writeapplications,andmoresophisticated electricalmodes(seebelow).Theequationforelectrostaticforcesbetweenaprobeanda surfacehavingdifferentpotentialsisgivenby: = −1/2(V 2 electrostaticPage 27 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes 2 Felectrostatic = −1/2(V ) dC dz (3.3) ItcanbeseenthatfromEquation3.3andEquation3.2thatthechangeinresonant frequencyisproportionaltothechangesincapacitanceasafunctionofthesecond derivativeofzspacing.Inotherwords,aslongasthereisanon‐zeropotentialbetween theprobeandsurface,thefrequency,andthustheamplitudeandphaseofoscillationwill besensitivetocapacityofthesurface. EFMhasbeenshowntodetecttrappedchargeonsurfaces[222],andinsomecases givesclearcontrastwherenoneisvisibleinthetopographysignal.However,ithasbeen reportedthatEFMispronetotopographicartefacts[223].EFM,likeMFMhasthegreat advantagethatitmaybecarriedoutwithastandardAFM.Asomewhatmore sophisticatedtechniquetomeasuretip–samplepotentialisscanningKelvinprobe microscopy(SKPM)[224,225].Figure3.23illustratestheportionoftheSKPM instrumentusedforequilibratingtheprobesurfacepotential.Theelectronicsusedfor mechanicallyvibratingthecantileverarenotshown. TheprincipleofoperationofSKPMissimple,thatiswhentwosurfaceshavethesame potentials,therewillbenoforcesbetweenthem,soinEquation3.3,ΔV=0.Toimplement thetechnique,aDCpotentialbias(VDC)isappliedtoaconductiveprobe,whichis furthermodulatedbyanACsignal(VAC),sothat Fig.3.23. Schematicillustrationofinstrumentalset‐upforscanning Kelvinprobemicroscopy. (p.76) Page 28 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes Fig.3.24. ExampleofKelvinprobeandelectricforcemicroscopy. AFMheightimage(A,shadedimage),Kelvinprobe(B),andEFM(C) imagesofcarbonnanotubesonagoldsurface.Theimagesarenotall inexactlythesameplace;theredarrowhighlightsaconnection betweentwonanotubesineachimage.Reproducedfrom[227],with permission. Vbias = VDC + VAC sin t (3.4) Inotherwords,theACvoltageisoscillatingattheresonantfrequencyofthecantilever [224].Thus,theprobe'selectricpotentialisvaryingatfrequencyω.Ifthesample's potentialisnotthesame,thedifferenceinelectricalpotentialwillcausethecantileverto mechanicallyvibrateatthefrequencyω,andwhichmeansthattheelectricalsignalfrom thephotodetectorwillbemodulatedatω.Afeedbackcircuitthencomparesωwithω mod,andoutputsaDCvoltagetothesamplethatminimizestheoscillationatωmod.This occurswhentheappliedpotentialVDC isequivalenttothesurfacepotentialVs .Sothe voltageVDC thatisrequiretominimizeωmodisdigitizedwiththeA/Dconverterand displayedonthePCasthepotentialimage[225,226].BySKPM,absolutevaluesofthe sampleworkfunctioncanbeobtainedifthetipisfirstcalibratedagainstareference sampleofknownworkfunction. 3.2.6ElectrochemicalAFM Althoughnotreallyaseparatemode,itisworthmentioningthatitisrathersimpleto studyasurfaceasafunctionofappliedpotentialusingtheAFM[228].Changesinsample topographywithappliedpotentialaretheresultsofelectrochemicalreactions,andsothis techniqueisknownaselectrochemicalforcemicroscopy.Insituimagingofsuch processesisachievedwithanelectrochemicalcellwhichisamodifiedliquidcellwiththe additionofelectrodestobiasthesampleandapotentiostat.Byrampingtheapplied potentialtotheoxidationorreductionpotentialofthesurfaceduringscanning,or betweenscans,itispossibletodirectlyobserveoxidationorreductionprocessesonthe samplesurface.Suchprocessestendtogiverisetosmall(orslow)changesinsample topography,hencetheusefulnessofelectrochemicalAFM.Furthermore,itispossible, usingmoremodificationsoftheinstrument,tocombineimagingwithelectrochemical measurementsatthenanoscale,atechniquereferredtoasscanningelectrochemical AFM[229].AnexampleimageshowingresultsfromelectrochemicalAFMisshownin Figure3.25. (p.77) Page 29 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes Fig.3.25. ElectrochemicalAFMexample.Imagesshowingthe morphologyofaCdTefilmduringelectrochemicaldepositionofAu, atvarioustimes(asshowninfigure)atapotentialof−0.35V. Reproducedwithpermissionfrom[230]. 3.2.7Thermalmodes ItispossibletousederivativesofAFMtomeasurethermalpropertiesofthesample [231].Typically,thisisdonebyusingaresistiveprobe,whichcanlocallyheatthesample ormeasurethetemperaturelocally,i.e.actasathermometer.Thefirstsuchprobes weretheso‐calledWollastonwireprobes,whichconsistofaveryfineplatinumwirebent intoav‐shape.Theapexofthevformedthetipoftheprobe.Later,micromachined probes,developedfromsiliconnitridecantilevers,withapalladiumlayerwhichthins greatlyatthetipapex,toactastheresistor,weredeveloped.Onecommonexperiment involvesapplyingapotentialtotheprobe,whichheatstheresistance.Asthesampleis scanned(incontactmode),heatfromtheprobewillflowintothesample,theamount dependingonthethermalpropertiesofthesample,andafeedbackcircuitadjuststhe currentflowingthroughtheresistor,tokeeptheresistance,andthusthetemperature, constant.Plottingthecurrentappliedtotheprobegivesthethermalimage,anda topographicalimageiscollectedsimultaneously.Anexampleofthesortofdatathatmay becollectedwiththistechniqueisshowninFigure3.26.Thismethodiscommonlytermed scanningthermalmicroscopy(SThM).ThethermalimageinSThMisthereforeamapof thermalconductivity,althoughitmightbenecessarytodeconvolvetopographic contributions[231].Byusingtemperaturemodulation(i.e.bysupplyinganACcurrentto theresistorratherthanaDCcurrent),thedepthsensitivitymaybechanged,allowingfor (p.78) Page 30 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes Fig.3.26. Exampleofscanningthermalmicroscopy.Thermal conductivityimageofasectionfromaglassfilament/cyanateresin composite.Theglassfibresclearlyshowgreaterthermal conductivitythanthepolymermatrix.Reproducedfrom[238]with permission. discriminationofburiedfeatures[232].Thismodealsoallowsfortheimagingofheat capacity[231].Inadditiontotheimaging‐typeexperiments,itispossibletoperformmany typicalthermalanalysisexperimentsusingasimilarset‐upsuchaslocalizedcalorimetry orthermo‐mechanicalanalysis[233–236].Theaimofallthesetechniquesisto characterizematerialsthermallyonthenanoscale.Assuchmostoftheseexperiments couldbeperformedmacroscopicallyonwholesamplesmuchmoreeasily,sothemain applicationisinheterogeneousmaterials.Aswellasspecializedprobes,SThMrequires somesimpleexternalcircuitry,andsoitsadoptionasastandardAFMtechniquehasnot beenwidespread.However,suchprobesarecommerciallyavailable,andthetechnique givesinformationnotavailablebyothermeans,soalargenumberofstudieshavebeen appliedtopolymercomposites[237–239];inaddition,micro‐organisms[231], pharmaceuticals[232,236,240],automotivecoatings[241],metalalloys[242]and electronicdevices[243]havebeenstudiedwithSThM.Theinterestedreaderisdirected toanexcellentreviewformoreinformationonthistechnique[231]. 3.3Surfacemodification Aswellasmeasuringsamplesurfaces,anAFMmaybeusedtomanipulateortomodify thesurfaces.Thefinecontroloftheprobemotionoverthesurfacemakesevena standardAFMaversatiletoolformanipulationsurfacesatthenanoscale.Therearea rangeoftechniquesthathavebeenusedtomodifysurfaces,notablyincludinglocal oxidation[244],scratching[245]anddip‐pennanolithography[246]. (p.79) UncontrolledsurfacemodificationisusuallyanundesiredfeatureofAFM,butit wasrealizedearlyinthehistoryofSPMthatwithcarethistechniquehadthepotentialto fabricatenanoscaledevices[247].Oneoftheearliestofthenanolithographictechniques tobedemonstratedwaslocaloxidation[248].Inthistechnique,abiasisappliedtothetip tocausecontactpotentialdifferencewhilescanningthesurface,resultingtypicallyinan oxidationofthematerialatthesamplesurface.Theseexperimentsarecommonlycarried Page 31 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes outonsiliconandresultinfeaturesofsiliconoxideatthesurface[244],althoughother oxidation‐initiatedreactionsarepossible[249,250].Asnotedpreviously,whenscanning incontactmode,aliquidmeniscuswillbepresentbetweenthetipandsamplesurface.In nano‐oxidationthismeniscusisvitalbecauseitprovidestheelectrolyteforoxidation. Becauseoftheimportanceoftheliquidbridgeforthereaction,theprocessisvery sensitivetohumidity,andthesizeofthemeniscushasbeenreportedasthefactor controllingthesmallestfeaturethatit'spossibletomanufacture[244].Localoxidationhas beenperformedincontact[251–253],intermittent‐contact[253],andnon‐contactmode [254].Ifthetipisinthenon‐contactregimewhenthebiasisapplied,acapillarylayercan spontaneouslyform,andithasbeensuggestedthatthewaterbridgeunderthese circumstancesissmallerthanincontactmode,leadingtosmallerwrittenfeatures[254]. Thistechniquehasalsobeenshowntobeapplicabletoparallelfabrication[255–257], whichisofgreatimportance,becausethemaindrawbackofAFM‐basednanolithography forfabricationisitsslowspeed[252].Still,whilelocaloxidationhasbeenusedtocreate nanoscopicfunctioningelectronicdevices[258,259],fabricationofindustriallyuseful structuresonalargescalebythistechniquehasyettobedemonstrated,evenusing parallelwritingtechniques. Tocarryoutsurfacemodificationwithscratchingtechniquesisaverysimpletechnique, andisoftenusedasaproofofprincipleexperimentforlithographyapplications,because itissimpletoapplytoarangeofmaterials.Structureshavebeenbuiltinpolymers, silicon,metalsandmorebyscratching[245,249].Inprinciple,allthatisrequiredisto applyahighnormalforcetothesample,andusethelithographiccontrolsintheAFM controlsoftwaretodirectthetipinthedesiredpattern.Inthisway,highlyintricate patternscanbeformedwiththistechnique.Unfortunately,unlikeoxidationorDPN,itis rarelyappliedtobuildstructureswithchemicallydifferentfeatures,sothenumberof usefulapplicationsisrelativelylow. Dip‐pennanolithographywasinventedin1999byMirkinandcoworkers[260],andhas beenshowntobeahighlyversatiletechnique.Thegreatadvantageofthistechniqueis thatalmostanymaterialthatcanbedepositedonasurfacecanbeusedandformedinto nanometre‐scalepatterns,althoughtypicallywater‐solublemoleculesorverysmall particlesareapplied[246].Theideaisanalogoustothatofamacroscopicpen.TheAFM tipisimmersed,ordippedintoasolutionofthemoleculetobegrafted.Withahydrophilic tip,andaqueoussolution,theAFMprobewillbecomecoatedinathinlayerofthewriting solution.Then,whenthetipisincontactwiththesubstrate,thegraftingmoleculesare appliedtothesurfaceviathewatercapillarylayer[260].Aschematicillustratingthisis showninFigure3.27. Likenano‐oxidation,thesizeofthewaterbridgeisacontrollingfactorinthedimensionof thewrittenfeatures,aswellassuchfactorsasset‐point,scanningspeed,diffusionofthe moleculesonthesurface,andtipradius[249,261].Examplesofthesortoffeaturesthat maybeproducedareshowninFigure3.28.Agreatvarietyof‘inks’havebeenused,and (p.80) Page 32 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes Fig.3.27. Schematicofdip‐pennanolithography,showinghowthe watermeniscusisusedtotransportmoleculestothesurface. Adaptedfrom[260]. patternshavebeencreatedfromorganicmolecules[260],proteins[262,263],synthetic peptides[264],DNA[246],polymers[263],inorganicnanoparticles[265]andmore[246, 249].Amajorapplicationofthissortoftechnologyisincreationofarraysofreceptorsfor paralleltesting,e.g.proteomics,genomics,etc.Forlargescaleparallelarraysofdiffering features,specializedDPNinstruments,ratherthancommercialAFMSaretypicallyused. Anumberofother,lesscommonlyusedmethodsexisttomodifysurfaceswithAFM [249,266].Theseincludethermomechanicalwriting,whichlikeSThMusesaresistancein theprobetocontrolthetemperatureatthetip[267].However,thetemperatureisused tomodifythesamplesurface,ratherthantoprobeit,andthehightemperatureis typicallyusedtomakeholesinpolymersurfaceswithoutriskofdamagingthetip.Thishas beeninvestigatedasahigh‐densitydatastoragetechnique,andviatheuseofparallel probes(theso‐called‘millipede’device[268]),hasbeenshowntobecapableof extremelyhighstoragedensity[269].SeveralauthorshavereportedtheuseoftheAFM todirectlymanipulateindividualparticles[270],molecules[271]andevenatoms[272, 273]onasurfacebyforexample,pushing,liftinganddroppingorcutting[249].These proceduresareinterestingforfundamentalstudiesbutaretooslowtobeofvalueas manufacturingtechniques.SomeexamplesofassemblyusingAFMareshowninSection 7.2.3.Finally,a Fig.3.28. ExamplesofAFM‐basedlithography.Left:polymeric patternsonsiliconformedbyanodicoxidation,showinglinewidths ofapproximately2nm.Reproducedwithpermissionfrom[250]. Centre:abit‐mapimageusedastheinputforadip‐pen nanolithography(DPN)routine.Right:AFM(lateralforce)imageof theresultingsurfacepatterns. (p.81) techniquecallednanograftingisavariantofdip‐pennanolithography[274,275].It hasthesameadvantageofflexibility–awidevarietyofmoleculesmaybeappliedtothe Page 33 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015 AFM modes surface[274,276].ThedifferenceisthatitinvolvesusingtheAFMtiptoremove moleculesfromapreviouslymodifiedsurface,sothatthemoleculesofinterest,whichare insolution,canformpatcheswithinthepreviouslayer[277].Thishastheadvantageof leavingthemoleculesofinterestsurroundedwithapotentiallyinertpassivatinglayer coveringthe(typically)metallicsubstrate,makingitusefulforexamplefabricationof devicesforbindingstudies[262]. Page 34 of 34 PRINTED FROM OXFORD SCHOLARSHIP ONLINE (www.oxfordscholarship.com). (c) Copyright Oxford University Press, 2014. All Rights Reserved. Under the terms of the licence agreement, an individual user may print out a PDF of a single chapter of a monograph in OSO for personal use (for details see http://www.oxfordscholarship.com/page/privacy-policy). Subscriber: UC Berkeley Library; date: 20 February 2015