Dynamic Light Scattering - Understanding the Basics

DynamicLightScatteringUnderstandingthe
Basics
SponsoredbyMalvernInstrumentsLtd Oct182013

TopicsCovered
Introduction
TheBasicsofDLS
TheBenefitsofDLS
LimitationsofDLS
DefiningFeaturesofDLSInstrumentation
AssessingtheExtras
Selection
Conclusion
AboutMalvernInstruments

Introduction
Dynamiclightscattering(DLS)isavaluedsizingtechniqueforproteins,colloidsand
dispersions,whichcomfortablyextendstothesub1nmregion.Theobservationof
scatteredlighthelpsdeterminedefiningcharacteristicsofaparticledispersionormolecular
solutionsuchasparticlesize,molecularweightandzetapotential.

ThisarticleexamplesthebenefitsandlimitationsofDLS,focusesonthoseaspectsof
systemdesignthatarecrucialindefiningperformance.

TheBasicsofDLS
SmallparticlesinadispersionorsolutionaresubjecttoBrownianmotion.DLSisdrivenby
collisionswiththesolventmoleculespresent,whichareinconstantmovementduetotheir
thermalenergy.ThespeedofBrownianmotioncanbedirectlymeasuredfromthe
scatteredlightpatternproducedbythemovingparticles,atechniqueknownasphoton
correlationspectroscopy(PCS)orquasielasticlightscattering(QELS)butpresently
referredtoasDLS.

TherelationshipbetweenthespeedofBrownianmotionofaparticleandthatparticles
sizeisdefinedbytheStokesEinsteinequation:

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Dynamic Light Scattering - Understanding the Basics

WhereD=Diffusionspeed,k=Boltzmannsconstant,T=absolutetemperature,=
viscosity,andDH=hydrodynamicradius.

Thisrelationshipshowshowsizecanbedeterminedfromdiffusionspeedprovidedthatthe
temperatureandcontinuousphaseviscosityofthesampleareknown.

TheBenefitsofDLS
ThebenefitsofDLSare:

Operatorscanattainusable,detaileddatawithoutneedingtohavesignificant
expertise.
Samplevolumesaresmall,downtojustafewmicroliters,makingthisanappealing
techniqueforearlystageresearchwherevaluablematerialsareinvolved.
DLSisessentiallyverygoodatmeasuringparticlesizeacrosstherange~0.1nmto
~10m.
Itsabilitytomeasuresmallparticlesisespeciallyvaluable,withmostsystemsgiving
accurate,reproducibledataforparticlesfrom2nmandabove.
Itispossibletomeasurebothdiluteandturbidsystemswiththeconcentrationrange
foranalysisreachingdownaslowas0.1ppmandupto40%w/v.
Thissizeandconcentrationrange,togetherwiththehighreproducibilityofthe
techniquemakesDLSsuitableforawiderangeofapplications.

LimitationsofDLS
ThelimitationsofDLSare:

Largeparticlesifpresentinevensmallquantitiesmaybeaccountedduringdata
analysis
SedimentationThisisparticularlylikelywithmoredenseparticles.Increasingthe
densityofthedispersantby,forexample,introducingsucroseisahelpfulstrategybut
onlyfordensitymatchinguptoaround1.05g/ml.
LowresolutionWhensizepopulationsarecloselyspaced,lessthanafactorof
threedifferenceinsize,DLSwillnotpreciselycharacterizeapolydispersesample,
makingitadvisabletoapplyseparationpriortomeasurement.

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Dynamic Light Scattering - Understanding the Basics

MultiplelightscatteringMultiplescatteringiswhenthescatteredlightfromone
particleisscatteredbyanotherbeforereachingthedetector,anditcompromisesthe
accuratecalculationofparticlesizeinmoreconcentratedsamples.
DispersantchoiceEventhoughmostdispersantsaresuitable,thosewitha
viscositygreaterthan100mPa.sinhibitreliablemeasurements.Furthermore,light
absorptionbythedispersantcaninterferewithdetection

DefiningFeaturesofDLSInstrumentation
Keyfeaturesoftheopticalsetupare:

ThelaserAstablelaserwithlownoisecharacteristics,asexemplifiedbycertain
HeliumNeongaslasers,isthemostsuitable.
TheopticalarrangementMeasuringonlyat90oenablestheproductionofasimple,
costeffectivesystemthatprovidesanappropriatelevelofsensitivityformany
applicationsasshowninFigure1.Forexperimentsthatrequirehighersensitivity
levelsorformoreconcentratedsamples,measuringatwideranglesispreferable.
Thedetector(s)Detectorsareoneoftwotypes:cheaper,lesssensitive
photomultipliersormoreexpensive,higherperformanceavalanchephotodiode
detectors(APD).

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Dynamic Light Scattering - Understanding the Basics

Figure1.ThekeyelementsofaDLSsystemarethelaser(1),measurementcell(2),
detector(3),attenuator(4),correlator(5)anddatahandlingPC(6).Detectorscanbe
placedateither90ooratawiderangleasexemplifiedbytheNIBSdetectorshownathere
at173o.

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Dynamic Light Scattering - Understanding the Basics

Figure2.Zetapotentialquantifiestherepulsionbetweenparticlesinasuspensionaiding
stabilitystudies

AnotheressentialrequirementforpreciseDLSmeasurementsisgoodtemperaturecontrol.
Builtintemperaturecontroliseasiertousethanattachinganexternalwaterbathsetup.
Inaddition,instrumentswithhighperformancecontrolsystemsmayofferfastsystem
warmupandtheabilitytorapidlychangetemperatureinordertostudytemperature
dependenteffectssuchasproteinmelting.

AssessingtheExtras
DLSisacceptedforparticlesizemeasurement,alsothecombinationofcomponentsthat
makeupahighlyspecifiedDLSsystemalsolendthemselvestothemeasurementof:

Absolutemolecularweight
Zetapotential
Proteincharge
Microrheology

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Dynamic Light Scattering - Understanding the Basics

Selection
Whileselectingtheinstrument,thesefactorsmustbeconsidered:

Whatismostimportant:speedoraccuracy?
Whatsamplesizeoneneedstomeasure?
Whattypeofsamplesaregoingtobemeasuredaretheytoxicforexample,or
especiallycorrosive?
Aretheoperatorslikelytobeexpertornoviceusers?Howmuchlightscattering
expertiseisavailable?

Conclusion
DLSisanestablishedmethodextensivelyusedformolecularandparticlesize
measurement,foratrulydiverserangeofsampletypes.Thismakesitimportanttoclosely
matchuserrequirementstothecapabilitiesofasystemwhilechoosinganinstrument.

Alongsideparticlesize,lightscatteringsystemsmayalsodeterminemolecularweight,
proteinchargeandzetapotential,orevenhavemicrorheologycapabilities.Thereisa
variationinsensitivityconsiderablyfromsystemtosystem,definingthesizeofparticlesor
moleculesthatcanbemeasuredrobustly,asdoestheabilitytomeasureathigh
concentrations.

AboutMalvernInstruments
Malvernprovidesthematerialsandbiophysicalcharacterizationtechnologyandexpertise
thatenablesscientistsandengineerstoinvestigate,understandandcontroltheproperties
ofdispersedsystems.

Thesesystemsrangefromproteinsandpolymersinsolution,particleandnanoparticle
suspensionsandemulsions,throughtospraysandaerosols,industrialbulkpowdersand
highconcentrationslurries.Usedatallstagesofresearch,developmentand
manufacturing,Malvernsinstrumentsprovidecriticalinformationthathelpsaccelerate
researchandproductdevelopment,enhanceandmaintainproductqualityandoptimize
processefficiency.

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Dynamic Light Scattering - Understanding the Basics

Thisinformationhasbeensourced,reviewedandadaptedfrommaterialsprovidedby
MalvernInstruments.

Formoreinformationonthissource,pleasevisitMalvernInstruments.

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Dynamic Light Scattering - Understanding the Basics

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