Anintroductiontocirculardichroismspectroscopy

Circulardichroism(CD)isthedifferenceintheabsorptionoflefthandedcircularlypolarised
light(LCPL)andrighthandedcircularlypolarisedlight(RCPL)andoccurswhenamolecule
containsoneormorechiralchromophores(lightabsorbinggroups).

Circulardichroism=A()=A()LCPLA()RCPL,whereisthewavelength

Circulardichroism(CD)spectroscopyisaspectroscopictechniquewheretheCDofmoleculesis
measuredoverarangeofwavelengths.CDspectroscopyisusedextensivelytostudychiral
moleculesofalltypesandsizes,butitisinthestudyoflargebiologicalmoleculeswhereit
findsitsmostimportantapplications.Aprimaryuseisinanalysingthesecondarystructureor
conformationofmacromolecules,particularlyproteinsassecondarystructureissensitivetoits
environment,temperatureorpH,circulardichroismcanbeusedtoobservehowsecondary
structurechangeswithenvironmentalconditionsoroninteractionwithothermolecules.
Structural,kineticandthermodynamicinformationaboutmacromoleculescanbederivedfrom
circulardichroismspectroscopy.

Measurementscarriedoutinthevisibleandultravioletregionoftheelectromagnetic
spectrummonitorelectronictransitions,and,ifthemoleculeunderstudycontainschiral
chromophoresthenoneCPLstatewillbeabsorbedtoagreaterextentthantheotherandthe
CDsignaloverthecorrespondingwavelengthswillbenonzero.Acirculardichroismsignalcan
bepositiveornegative,dependingonwhetherLCPLisabsorbedtoagreaterextentthanR
CPL(CDsignalpositive)ortoalesserextent(CDsignalnegative).Anexamplecircular
dichroismspectrumofasamplewithmultipleCDpeaksisshownbelow,demonstratinghow
CDvariesasafunctionofwavelength,andthataCDspectrummayexhibitbothpositiveand
negativepeaks.

Circulardichroismspectraaremeasuredusingacirculardichroismspectrometer,suchasthe
Chirascan,whichisahighlyspecialisedderivativeofanordinaryabsorptionspectrometer.CD
spectrometersmeasurealternatelytheabsorptionofLandRCPL,usuallyatafrequencyof
50kHz,andthencalculatethecirculardichroismsignal.

1. Understanding Circular Dichroism

Thebasicsofpolarisation

Toreallyunderstandcirculardichroism,onemustfirstunderstandthebasicsofpolarisation.
Linearlypolarisedlightislightwhoseoscillationsareconfinedtoasingleplane.Allpolarised
lightstatescanbedescribedasasumoftwolinearlypolarisedstatesatrightanglestoeach
other,usuallyreferencedtotheviewerasverticallyandhorizontallypolarisedlight.

Vertically Polarised Light

Horizontally Polarised Light

Ifforinstancewetakehorizontallyandverticallypolarisedlightwavesofequalamplitudethat
areinphasewitheachother,theresultantlightwave(blue)islinearlypolarisedat45degrees.
 45 Degree Polarised Light

Ifthetwopolarisationstatesareoutofphase,theresultantwaveceasestobelinearly
polarised.Forexample,ifoneofthepolarisedstatesisoutofphasewiththeotherbya
quarterwave,theresultantwillbeahelixandisknownascircularlypolarisedlight(CPL).The
helicescanbeeitherrighthanded(RCPL)orlefthanded(LCPL)andarenonsuperimposable
mirrorimages.

Theopticalelementthatconvertsbetweenlinearlypolarisedlightandcircularlypolarisedlight
istermedaquarterwaveplate.Aquarterwaveplateisbirefringent,i.e.therefractiveindices
seenbyhorizontallyandverticallypolarisedlightaredifferent.Asuitablyorientedplatewill
convertlinearlypolarisedlightintocircularlypolarisedlightbyslowingoneofthelinear
componentsofthebeamwithrespecttotheothersothattheyareonequarterwaveoutof
phase.ThiswillproduceabeamofeitherleftorrightCPL.

Left Circularly Polarised (LCP) Light

 Right Circularly Polarised (RCP) Light

Thedifferenceinabsorbanceoflefthandandrighthandcircularlypolarisedlightisthebasis
ofcirculardichroism.AmoleculethatabsorbsLCPandRCPdifferentlyisopticallyactive,or
chiral.

2. Chiral Molecules

Theoriginofopticalactivity

Inthefirstsectionofthistutorial,westatedthatcirculardichroismisthedifferential
absorbanceoflefthandcircularlypolarisedandrighthandcircularlypolarisedlight,andthat
chiralmoleculesexhibitcirculardichroism.Butwhatarechiralmolecules?

Chiralmoleculesexistaspairsofmirrorimageisomers.Thesemirrorimageisomersarenot
superimposableandareknownasenantiomers.Thephysicalandchemicalpropertiesofapair
ofenantiomersareidenticalwithtwoexceptions:thewaythattheyinteractwithpolarised
lightandthewaythattheyinteractwithotherchiralmolecules.

Circularbirefringenceandopticalrotation

Chiralmoleculesexhibitcircularbirefringence,whichmeansthatasolutionofachiral
substancepresentsananisotropicmediumthroughwhichleftcircularlypolarised(LCPL)and
rightcircularlypolarised(RCPL)propagateatdifferentspeeds.Alinearlypolarisedwavecan
bethoughtofastheresultantofthesuperpositionoftwocircularlypolarisedwaves,oneleft
circularlypolarised,theotherrightcircularlypolarised.Ontraversingthecircularlybirefringent
medium,thephaserelationshipbetweenthecircularlypolarisedwaveschangesandthe
resultantlinearlypolarisedwaverotates.Thisistheoriginofthephenomenonknownas
opticalrotation,whichismeasuredusingapolarimeter.Measuringopticalrotationasa
functionofwavelengthistermedopticalrotatorydispersion(ORD)spectroscopy.

Circular birefringence - the orange cuboid represents the sample

Circulardichroism

Unlikeopticalrotation,circulardichroismonlyoccursatwavelengthsoflightthatcanbe
absorbedbyachiralmolecule.AtthesewavelengthsLeftandrightcircularlypolarisedlight
willbeabsorbedtodifferentextents.Forinstance,achiralchromophoremayabsorb90%ofR
CPLand88%ofLCPL.Thiseffectiscalledcirculardichroismandisthedifferenceinabsorption
ofLCPLandRCPL.Circulardichroismmeasuredasafunctionofwavelengthistermedcircular
dichroism(CD)spectroscopyandistheprimaryspectroscopicpropertymeasuredbyacircular
dichroismspectrometersuchastheChirascan.

Circular dichroism - the orange cuboid represents the sample

Opticalrotationandcirculardichroismstemfromthesamequantummechanicalphenomena
andonecanbederivedmathematicallyfromtheotherifallspectralinformationisprovided.
Therelationshipbetweenopticalrotatorydispersion,circulardichroism,absorptionspectra
andchiralityareshownbelow,withacomparisonofthetwoenantiomersofcamphor
sulphonicacid.

CD, ORD and Absorbance spectra of R and S forms of camphor sulphonic acid

AlthoughORDspectraandCDspectracantheoreticallyprovideequivalentinformation,each
techniquehasbeenusedforverydistinctapplications.Opticalrotationatasinglewavelength
isusedasageneralmeasurementtoolforchiralmolecules,todetermineconcentrationandas
adeterminantofchiralpuritycomparedtoaknownstandard.Thesimplicityandlowcostof
theexperimentandinstrumentationmakesitidealforthisapplication.Circulardichroism
spectraontheotherhandarebetterspectrallyresolvedthanORDspectra,andconsequently
moresuitableforadvancedspectralanalysis.

3. Chirality and Biology

Circulardichroismandthestudyofbiologicalmolecules

Circulardichroismisaconsequenceoftheinteractionofpolarisedlightwithchiralmolecules.
Thevastmajorityofbiologicalmoleculesarechiral.Forinstance,19ofthe20commonamino
acidsthatformproteinsarethemselveschiral,asareahostofotherbiologicallyimportant
molecules,togetherwiththehigherstructuresofproteins,DNAandRNA.Thehighlychiral
chemistryofbiologicalmoleculeslendsitselfwelltoanalysisbycirculardichroismandthe
studyofbiologicalmoleculesisthemainapplicationofthetechnique.

Alargesubsetoftheuseofcirculardichroisminbiochemistryisintheunderstandingofthe
higherorderstructuresofchiralmacromoleculessuchasproteinsandDNA.Thereasonforthis
isthattheCDspectrumofaproteinorDNAmoleculeisnotasumoftheCDspectraofthe
individualresiduesorbases,butisgreatlyinfluencedbythe3dimensionstructureofthe
macromoleculeitself.Eachstructurehasaspecificcirculardichroismsignature,andthiscanbe
usedtoidentifystructuralelementsandtofollowchangesinthestructureofchiral
macromolecules.

Themostwidelystudiedcirculardichroismsignaturesarethevarioussecondarystructural
elementsofproteinssuchasthehelixandthesheet.Thisisunderstoodtothepointthat
CDspectrainthefarUV(below260nm)canbeusedtopredictthepercentagesofeach
secondarystructuralelementinthestructureofaprotein.Someofthecommonprotein
secondarystructuralelementsandtheCDspectraassociatedwiththemareshownbelow:

The secondary structure conformation and the CD spectra of protein structural elements. Right is an
example of the backbone conformation of a peptide in an -helix and left is the conformation of a peptide
in a -sheet. In the centre are the associated CD spectra for these different conformations.

Therearemanyalgorithmsdesignedforfittingthecirculardichroismspectraofproteinsto
provideestimatesofsecondarystructure.TheproteinsecondarystructureCDanalysis
softwaredistributedwiththeChirascanisCDNN.

Secondarystructurepredictionisonlypartofthepowerofcirculardichroismspectroscopy.
Changesincirculardichroismspectraareverygoodproxiesforchangesinthestructureofa
molecule.Couplethiswiththefactsthat(i)spectracanberecordedinminutesand(ii)single
wavelengthkineticscanberecordedfrommillisecondsonwards,CDisaparticularlypowerful
tooltofollowdynamicchangesinproteinstructure.Forinstancechangesinducedbychanging
temperature,pH,ligands,ordenaturantsareallcommonlyused.

Apowerfulapplicationofcirculardichroismistocomparetwomacromolecules,orthesame
moleculeunderdifferentconditions,anddetermineiftheyhaveasimilarstructure.Thiscan
beusedsimplytoascertainifanewlypurifiedproteiniscorrectlyfolded,determineifa
mutantproteinhasfoldedcorrectlyincomparisontothewildtype,orfortheanalysisof
biopharmaceuticalproductstoconfirmthattheyarestillinacorrectlyfoldedactive
conformation.

4. CD Spectrometer Operating Principles

Allthedetailsforafullunderstanding

Circulardichroism(CD)isthedifferenceinlightabsorbancebetweenleft(LCPL)andright
circularlypolarisedlight(RCPL)andcirculardichroismspectrometers(spectrophotometers)
arehighlyspecialisedvariationsoftheabsorbancespectrophotometer.

Acirculardichroismspectrophotometerisalsocommonlytermedacirculardichroism
spectropolarimeteroracirculardichrograph.Mostmoderncirculardichroisminstruments
operateonthesameprinciples,whichisdemonstratedintheslideshowatthebottomofthe
page.Thereisasourceofmonochromaticlinearlypolarisedlightwhichcanbeturnedinto
eitherleftorrightcircularlypolarisedlightbypassingitthroughaquarterwaveplatewhose
uniqueaxisisat45degreestothelinearpolarisationplaneasdescribedinthesectionabout
polarisedlight.

Insteadofastaticquarterwaveplate,acirculardichroismspectrophotometerhasa
specialisedopticalelementcalledaphotoelasticmodulator(PEM).Thisisapiezoelectric
elementcementedtoablockoffusedsilica.Atrest,whenthepiezoelectricelementisnot
oscillating,thesilicablockisnotbirefringent;whendriven,thepiezoelectricelementoscillates
atitsresonancefrequency(typicallyaround50kHz),andinducesstressinthesilicainsucha
waythatitbecomesbirefringent.Thealternatingstressturnsthefusedsilicaelementintoa
dynamicquarterwaveplate,retardingfirstverticalwithrespecttohorizontalcomponentsof
theincidentlinearlypolarisedlightbyaquarterwaveandthenviceversa,producingleftand
thenrightcircularlypolarisedlightatthedrivefrequency.Theamplitudeoftheoscillationis
tunedsothattheretardationisappropriateforthewavelengthoflightpassingthroughthe
silicablock.

Ontheothersideofthesamplepositionthereisalightdetector.Whenthereisnocircularly
dichroicsampleinthelightpath,thelighthittingthedetectorisconstant.Ifthereisacircularly
dichroicsampleinthelightpath,therecordedlightintensitywillbedifferentforrightand
leftCPL.UsingalockinamplifiertunedtothefrequencyofthePEM,itispossibletomeasure
thedifferenceinintensitybetweenthetwocircularpolarisations(vAC).Theaveragetotallight
intensityacrossmanyPEMoscillations(vDC)canbeusedtoscalethesizeofthelockin
amplifiersignaltotakeintoaccountvariationsintotallightlevel.Bothsignalscanberecorded
andfromthemthecirculardichroismsignalcanbecalculatedeasilybydividingthevAC
componentbythevDCsignal.

Gisacalibrationscalingfactortoprovideeitherellipticityordifferentialabsorbance.The
sectionaboutCDUnitsandtheirinterconversionexplainshowellipticityanddifferential
absorbancearerelated.

5. CD Spectrometer Performance

Thedesignanditseffectonoperation

Thelimitofdetectionofacirculardichroism(CD)spectrophotometer(orany
spectrophotometer)isdeterminedbyitssignaltonoise(S/N)characteristics:thebetterthe
S/N,thebetteritslimitofdetection.

Thesignaltonoiseratioislimitedbyphotonshotnoise,whichisthestatisticalvariationabout
anaverageinthenumberofphotonsperunittimedetectedbythelightdetector.The
quantisednatureofphotonsandtheirrandomarrivalatthedetectormeansthatalthoughthe
averagenumberofphotonsdetectedpersecondmaybesay5,thenumberinanyparticular
onesecondintervalmaybe0,2,7orsomeothernumber.Thus,ameasurementmustbe
madeoverasufficientlylongperiodoftimetodeterminethetrueaverageandthetimetaken
todeterminethetrueaveragewillbeinverselyproportionaltoS/N.Itisthereforeimportantto
designacirculardichroismspectrophotometertomaximiseitsS/Ncharacteristics.
Ageneralrelationshipbetweenthecontributingfactorstothesignaltonoiseinanoptical
spectrometercanbewrittenas:

whereQ=detectorquantumefficiency,I=lightintensity,t=timescaleofthemeasurement.

Fromthisitisapparenttherearethreewaystoimprovethesignaltonoiseofacircular
dichroismspectrophotometer:increasetheintensityoftheincidentlinearlypolarised
monochromaticlight,increasethequantumefficiencyofthedetector,orspendmoretime
collectingandaveragingdatapoints.

Thefirsttwofactors,lightintensityanddetectorperformance,arethosethatcanbe
influencedbythedesignofacirculardichroismspectrophotometerandworktogetherto
lowerthelastfactor,thetimerequiredtocarryoutameasurement.Thehigherthelight
throughputandbetterthedetectorefficiency,thelesstimeittakestocollectqualitydataor,
equally,thehigherthequalityofdatathatcanbecollectedintimelimitedexperimentssuch
asstoppedflowmeasurements.

Increasingtheintensityoftheincidentlightisthemainavenueforincreasingtheperformance
ofcirculardichroismspectrophotometersandthisfindsitsultimateexpressionintheuseof
synchrotronlightsourcesforCDspectroscopy.Synchrotronfacilitiesprovidetremendouslight
intensityacrossaverywidespectralrangeofwavelengthsbutaccesstothemisexpensiveand
limitedandtheiruseisrestrictedtothemorecuttingedgeapplicationsofcirculardichroism.
ForthevastmajorityofCDexperiments,ahighintensitybenchtopsourceistheonlypractical
option:AppliedPhotophysicsChirascanhasbeendesignedfromthegrounduptomaximise
thelightthroughputfromitsXearclampsourcetothesample.

TheChirascanmonochromatorusestwosynthetic,singlecrystalquartzprismsinsteadofthe
diffractiongratingsthatmostpeoplearefamiliarwithfromnormalabsorbance
spectrophotometers.Quartzprismsaremoreefficientthandiffractiongratingsforaverywide
rangeofwavelengths,particularlyintheUV.Quartzisalsobirefringentandtheprismsnotonly
disperselightintothecomponentwavelengthsbutalso,becauseoftheirbirefringence,
dispersethelinearlypolarisedcomponents,oneofwhichisselectedforconversionto
circularlypolarisedlight.Afurtheradvantageofprismsistheydonotpasssecondorder
multiplesofthedesiredwavelength,whichisamajorsourceofstraylightingratingbased
monochromators.

Unlikethedispersionofagrating,whichislinearandhighlycustomisable,thedispersionofa
prismisnonlinearandissetbythepropertiesoftheprismmaterial.Consequentlytheoptics
andmechanicsofaprismmonochromatorhavetobemorecomplexthanagrating
monochromator,withtheneedtoconstantlyvarytheslitwidthasafunctionofwavelengthto
maintainaconstantbandpass,andacomplexrelationshipbetweenprismmovementand
wavelength.However,thelargewavelengthdispersionintheUVmeansthatwiderslitscanbe
usedevenatasmallspectralbandpass,whichmeansgreaterlightcollectionefficiency
throughouttheUVregion.Thelargemajorityofcirculardichroismapplicationsarecarriedout
intheUVanditisatthesewavelengthsthatthecharacteristicsofaprismareamajor
advantage.
Inadditiontothehighintensitytoimprovesignaltonoise,thelightfromthemonochromator
musthaveaverylowstraylightcontentandaveryhighpurityoflinearpolarisationtoprovide
accuratemeasurements.Allofthesethreekeyelementshavebeenoptimisedinthe
Chirascancirculardichroismspectrophotometerandhavebeenachievedbykeydesign
featuresoftheChirascanmonochromator.

ThesecondinfluenceontheS/Nisthequantumefficiencyofthedetectorused.i.e.its
efficiencyinturninganincidentphotonintoanelectronicsignal.Incirculardichroism
spectrophotometers,thedetectorofchoiceinthelastfewdecadeshasbeenthe
photomultipliertube.Thequantumefficiencyofthesedetectors,whicharetraditionallyused
inspectroscopyforUVandvisiblelightdetection,hasremainedfairlystaticoverthatperiod.
Recently,advancesinphotodiodetechnologyhaveresultedinnew,highgain,largeareasolid
statedetectors,whichprovidesignificantimprovementsinquantumefficiencyintheultra
violet,visibleandnearinfraredregionswhencomparedwithphotomultipliertubes(see
Figure1).Oneofthesenewhighperformancesolidstatedetectorsisfeaturedinthenew
ChirascanplusanditgivesfurtherS/Nimprovementsoverthephotomultiplierbased
Chirascanspectrometer.Thequantumefficiencyimprovementoutlinedbelowresultsin
significantsignaltonoiseimprovementsoverandabovethealreadyhighperformanceofthe
standardChirascan.

Figure 1. Quantum efficiency of Chirascan Plus detector vs a standard photomultiplier tube

 6. CD Signatures of Structural Elements

Circulardichroismsignaturesofsecondarystructuralelements
 7. CD Units & Conversions

Allrelationshipsexplained

Circulardichroism(CD)isusuallyunderstoodandactuallymeasuredasthedifferential
absorbanceofleft(ALCP)andrightcircularlypolarised(ARCP)light,andsocanbeexpressed
as:

A=ALCPARCP

Takingintoaccountcellpathlengthandcompoundconcentration,wecanarriveatamolar
circulardichroism().

=LCPRCP=A/(Cxl)

WhereLCPandRCParethemolarextinctioncoefficientsforLCPandRCPlightrespectively,
C=molarconcentration,andl=pathlengthincentimeters.

AnotherimportantunitismeanresiduemolarcirculardichroismMR.Thisisaunitspecific
forproteins,andreportsthemolarcirculardichroismforindividualproteinresiduesinsteadof
wholeproteinmolecules.Thisallowseasycomparisonofdifferentproteinswithvastly
differentmoleculeweights.Therearetwowaystocalculatethisdependingonhowmuch
informationisknownabouttheprotein.

Theconcentrationofprotein(C)inmolarismultipliedbythenumberofaminoacids(N)inthe
proteintoprovidethemeanresidueconcentration(CMR):

CMR=CxN

MR=A/(CMRxl)

AnestimatecanbedeterminedforCMRifthesequenceoftheproteinisn'tknown,usingthe
averageaminoacidresidueweightof113daltons,andtheconcentrationofprotein(P)ingL1

CMR=P/113

Aandarethemostintuitiveunitsformanybiochemists,astheyarederivedfromthe
familiarconceptofUV/Visabsorbancespectroscopy,anditisalsohowmodernCDinstruments
actuallymeasurecirculardichroism.CDcanalsobeexpressedasdegreesofellipticity(),
whichisalegacyofpolarimetry,andsuchunitsarefrequentlyusedintheliterature.Inthe
contextofmodernCDspectroscopy,theseunitsarearchaicandcanbeconfusing.The
relationshipbetweenAandareexplainedbelow.

ThedescriptionofellipticityissomewhatmorecomplexthanA.Linearlypolarisedlightwhen
passedthroughacirculardichroicsamplewillbecomeellipticallypolarised.Elliptically
polarisedlightislightthatisnotfullycircularpolarised,butinsteadisellipticalinshape.Thisis
becausethecircularpolarisedcomponentsoftheoriginallinearpolarisedlightarenownotof
equalmagnitudesduetodifferentialabsorbance(circulardichroism).Theeffectis
demonstratedbelow:
 Circular dichroism as ellipticity - the orange cuboid represents the sample

Thedegreeofellipticity()isdefinedasthetangentoftheratiooftheminortomajorelliptical
axis,andisillustratedbelow:

Linear polarised light has 0 degrees of ellipticity (), while fully LCP or RCP will have + or - 45 degrees
respectively.
Theadvantageofcirculardichroismellipticityasameasurementunitisthatitismoreeasily
relatedtoopticalrotationmeasurementsandpolarimetry.Bothellipticityandopticalrotation
aremeasurementsofchangesinpolarisationstateofalinearpolarisedanalyzerbeam,and
bothhavethesameunitsandsimilaramplitudesforagivensample.Thissimilarityaidsin
comparisonofopticalrotationandcirculardichroismmeasurements,ausefulabilitywhen
circulardichroismspectroscopyfirststartedtobewidelyused,backinthe1960's.

FortunatelyitisveryeasytointerconvertbetweenandA:

A=/32.982

Note:Duetothesmallsizeofmanymeasurements,isoftenquotedasmillidegrees(m)or
1/1000ofadegree.

MolarellipticitycanbemanipulatedinthesamewayasA.Forinstancetakingintoaccount
concentrationandcellpathlengthaccordingtoBeerLambertslaw,wecanderivea
measurementofmolarellipticity[].Followingpolarimetricconventions,molarellipticityis
reportedindegreescm2dmol1,ordegreesM1m1whichareequivalentunitsasshown
below.

Molarellipticitycanbecalculatedusingthefollowingequation:

[]=100x/(Cxl)

Cistheconcentrationinmolar,andlthecellpathlengthincm.Thefactorof100convertsto
pathlengthinmeters.

MolarCirculardichroismandmolarellipticitycanbeconverteddirectlyby:

=[]/3298.2

Thisfactorisahundredfoldlargerthanbetweenrawabsorbanceandellipticityduetothe
conversionbetweenmolarextinctiondefiningpathlengthsincentimetersandellipticityhaving
pathlengthdefinedinmeters.

Anotherimportantunitismeanresidueellipticity[]MR.Thisisaunitspecificforproteins,and
reportsthemolarellipticityforindividualproteinresiduesinsteadofwholeproteinmolecules.
Thisallowseasycomparisonofdifferentproteinswithvastlydifferentmoleculeweights.There
aretwowaystocalculatethisdependingonhowmuchinformationisknownaboutthe
protein.

Theconcentrationorprotein(C)inmolarismultipliedbythenumberofaminoacids(N)inthe
proteintoprovidethemeanresidueconcentration(CMR):

CMR=CxN

[]MR=100x/(CMRxl)
AnestimatecanbedeterminedforCMRifthesequenceoftheproteinisn'tknown,usingthe
averageaminoacidresidueweightof113daltons,andtheconcentrationofprotein(P)ingL1

CMR=P/113

FortunatelytheProDatasoftwareforChirascancanconverteasilybetweenalltheseunits,
withaminimumofuserintervention.

WeatAppliedPhotophysicshopethatyouhavefoundthetutorialincirculardichroisma
usefulresource.IfyouwanttofindoutmoreaboutAppliedPhotophysics'rangeofcircular
dichroismspectrometers,thenpleasecontactsales@photophysics.com.