WT brochure spr - Metrohm

THE INSTRUMENTSTHE POSSIBILITIESTHE SOFTWARESurfacePlasmonResonanceInstrumentsDEDICATEDTO RESEARCH

THEINSTRUMENTS- Unique possibilities -Autolab ESPRIT allowsthe user to combineSPR measurements withelectrochemical measurementsAUTOLAB ESPRITThe surface plasmon resonancetechnique helps the scientist to analysebi-molecular interactions in real time,without labeling requirements.The study and characterization ofmolecular interactions are essential toexplore the structure-function relationshipsof biomolecules and to aid ourunderstanding of biological systems inlife sciences.Combining electrochemistry withSPR makes the Autolab ESPRIT perfectfor biosensor and DNA/RNA or proteinchip research.The double channel ESPRIT isdesigned with an open cuvette system,giving the user access to the cuvette aswell as the gold disk.The first channel is used to measurethe specific binding of the sample, thesecond channel is used to compensatefor non-specific binding, matrix andtemperature effects.The integrated high throughputautosampler provides the possibility tomeasure multiple samples automaticallywithout attendance, it also provides theoption to recover precious samples orunknown interacted molecules.ESPRIT comes with powerful dataacquisition software, which includesautomation tools for instrument control.All recorded data is evaluated in aseparate kinetic evaluation package, theraw data can also be exported into anythird party file formats. An additionalsoftware component to comply with GLPguidelines is available.Application areas● Chemical sensors● Affinity sensors● ImmunosensorsFundamental prerequisites● Efficient and stable immobilization● Complete retention of the biologicalproperties of the sample under study● Accessibility of biomoleculesKey features● Automated high throughput● Open cuvette system with referencechannel● Large dynamic refractive index rangePAGE 3

THE INSTRUMENTS- Compact SPR -Entry levelsingle channelSPR instrument withESPR capabilitiesAUTOLAB SPRINGLESPRINGLE is the latest addition tothe Autolab family of instruments. It isa single channel surface plasmon resonanceinstrument, based on the opticalprinciple of the ESPRIT.The compact design makesSPRINGLE an affordable alternative forelectrochemists who want to acquire anSPR signal during electrochemicalmeasurements or for biochemists whowant to get acquainted with the SPRtechnique in general.To enable electrochemicalmeasurements, the Autolab SPRINGLEcan be connected to any of our Autolabpotentiostat/galvanostat instruments.The SPR data can be used as a referenceor to complement results of electrochemicalexperiments.For example, during electropolymerization,SPR can be used tomonitor the growth and properties ofthe polymer layer on the surface.SPRINGLE uses disposable pipettetips to aspirate and dispense samples,allowing fast and clean sample changing.The inject sequence is semi-automatic:the only manual action is placinga sample vial under the pipette tip, thesample is aspirated and injected in thecuvette when measurement starts. It isalso possible to manually inject a sampleusing a pipettor.The open cuvette system allowsquick access, easy electrochemistry anddoes not have any clogging or congestionproblems, the open system also allowsmultiple sample additions.Application areas● Chemical sensors● Affinity sensors● ImmunosensorsFundamental prerequisites● Efficient and stable immobilization● Complete retention of the biologicalproperties of the sample under study● Accessibility of biomoleculesKey features● Affordable compact instrument● Open cuvette system● Large dynamic refractive index range● Single channelPAGE 4

THE TECHNIQUE- Maximum flexibility -THE SPR PRINCIPLESurface plasmon resonance occurswhen p-polarized light hits a gold layerat a certain angle under conditions oftotal internal reflection. An evanescentwave is generated and the energy isabsorbed by the free electrons in the goldlayer, generating resonance plasmonsand causing a reduction in the intensityof the reflected laser light.Since molecules are immobilizedon the gold surface, the refractive indexchanges, altering the angle at which theplasmon effect occurs.Optical unitThe Autolab SPR instruments usea scanning mirror to alter the angle ofincidence of the laser light. The mirrorscans 4 degrees at a certain offset angle,with a frequency of 76 Hz.The minimum in reflectionintensity, known as SPR dip, is detectedautomatically and plotted as a relativeSPR angle in the software. The offsetangle can be manually controlled witha spindle, providing a large dynamicrange.The high scan frequency results inlow noise measurements. The laser spothas an area of 2 mm 2 on the sensorsurface. Measured data will be anaverage of all events in that area.This has the advantage that anyinhomogeneity in, or defects of any ofthe additional layers on top of the goldsurface will be eliminated.SubstratesEco Chemie provides standardsensor disks: bare gold substrates madeof BK-7 glass with 48 nm of golddeposited on a thin layer of titanium.The low cost sensor disks areplaced on a retractable slider with ahemicylinder (also BK-7). To increasecompatibility, a special slider formeasurements on Biacore sensorchipsis available.The range in refractive indices ofthe sample solution under study canbe extended by using different types ofglass hemicylinders and substrates.CuvetteExcept for the diameter of thecuvette and the geometry of the needle,the following parameters can becontrolled by the user:● Mixing volume● Speed or frequency of mixing● Distance of the pipette tip/needle tothe sensor surface● Volume of the solution in the cuvette● Viscosity of the solution● Temperature of the solutionThis makes the instrument verysuitable for fundamental research. It isalso possible to use custom-builtcuvettes and/or cuvettes of differentmaterials.The ESPRIT and SPRINGLE havea unique electrochemical cuvette thatallows measurements on a 3-electrodesystem;● Fixed contact point tothe gold layer of thesensor disk (workingelectrode)● Reference electrode,which is replaceable● Counter electrode isa fixedPlatinumelectrodeLiquid handling unitSample handling can be donemanually, semi-automatic or fully automatic.In the Autolab ESPRIT, the twochannels of the cuvette use independentsyringe pumps to assure reproducibleexperimental conditions.Fully automaticThe autosampler is controlled bythe liquid handling tool in the software.Samples are taken from the microliterplate and injected in the cuvette. Theautomatic control allows the user to runmeasurements continuously.The software has predefined rackcodesfor different kinds of microliterplates, but it is also possible to definecustom-made sample holders.Semi-automatic controlThe sample is aspirated in theneedle/pipette tip or pipetted in thecuvette manually.Other liquid handling functionalitylike dispensing, draining, washing,mixing of sample and measurement isperformed automatically.Manual controlAfter the sample is pipetted intothe cuvette, all following steps likemixing, draining, washing, starting ofmeasurement, rinsing etc. are controlledmanually from the software with asimple mouse click.Autolab SpincoaterThe Autolab Spincoater is anaffordable tabletop spin coating systemthat can be used to coat uniform layersof for example polymers or metal oxideson a substrate.The system has a bowl with adiameter of 13.5 cm. Rotation speed canbe controlled manually from 100 - 10,000rpm with an accuracy of 2 rpm in stepsof 100 rpm.SPECIFICATIONS AUTOLAB SPINCOATER● Standard sample size● Motor speed range● Manual speed setting● Acceleration/deceleration25 mm ∅ disk100 - 10,000 RPM100 - 10,000 RPM in 100 RPM steps4,000 RPM/sPAGE 5

THE SOFTWARE- Easy to use -The sophisticatedcontrol and dataacquisition softwaremakes complextasks easyTHE AUTOLAB SPR SOFTWAREThe Autolab SPR software packagecontains a program for data acquisitionand a separate program for kineticevaluation. Both programs operate underWindows 2000 and Windows XP.Data acquisitionThe data acquisition softwarecontains all necessary tools to controlthe instrument and to measure and storedata of interaction measurements.After stabilizing the sensor surface,phase one in interaction measurementsis to make the sensor surface biospecificby immobilizing a ligand, the nextphase is the measurement of analytes onthe biospecific surface, and finally thesensor surface is regenerated.All four measurement phases(baseline, association, dissociation andregeneration) are plotted in the ‘DataPresentation’ window and are controlledthrough measurement sequences.SequencerA sequence is a list of commandsthat is used to automate the control ofthe instrument.The Autolab SPR software comeswith a complete set of sequences for allkinds of measurements. It is alsopossible to create new sequences or toedit existing ones in the ‘Sequencer’window.The sequencer, combined with theautosampler functionality of the Esprit,allows you to operate the instrument 24hours per day, 7 days per week.Multiple sample injections areexecuted by the sequencer based onsample positions indicated in the ‘AutosamplerControl’ window.In this window, sample positions,identification and incubation times canbe specified.Sample recoveryIt is possible to add samplesduring measurement, to perform duplomeasurements on each sample and toperform sample recovery.Data PresentationThe 'Data Presentation' windowshows all measured data; the baseline,the association, dissociation andregeneration phase. The temperature ofthe slider is also recorded at the sameinterval, it can be used to compensate fortemperature effects.Users can add markers to any datapoint on the plot and add comments onspecial events or specific areas ofinterest that are logged into an eventrecorder. The double channel ESPRITinstrument has the option to plot thedata of both channels as well as thedifferential signal.Measurement integrityThe data acquisition software wasdeveloped to meet GLP guidelinesand facilitates compliance with 21 CFRPart 11. The software security featuresinclude:● Log in with user ID and password● Multiple access levels● Protected sequences for standard use● Protection of data with electronicsignatures● Log and audit trailsKinetic evaluationThe SPR data collected by the dataacquisition software can be analysedwith the kinetic evaluation software.With the kinetic evaluationsoftware the following parameters canbe calculated:● Kinetic rate constants● Affinity constants● Thermodynamic parametersThe above parameters can becalculated using the multiple curvesmeasured with different concentrations.The actual curves can be fitted todifferent kinetic models.PAGE 6

TECHNICAL SPECIFICATIONSSPECIFICATIONSESPRITSPRINGLE● Measuring principle● Transducer principle● Liquid handling● Parallel channels● Fixed wavelength● Sample loading and injection● Mixing● Autosampler● Pumps● Flow rate range● Sample volume● Offset of SPR angle by spindle● Dynamic range● Angle resolution● Refractive index● Refractive index resolution● Minimum molecular weight● Measuring frequency● Time interval range● Baseline noise● Sensor disk● Standard supplied cuvette● Additional cuvette● Biacore sensor chip adaptor● Weight● Dimensions (W × D × H)● Power requirementsSurface plasmon resonanceScanning mirrorCuvette system2670 nmManual, semi-automatic and automaticcontrolContinuous with autosampler needlesIntegratedSyringe and peristaltic0.8 - 227 µl/s20 - 150 µl62˚ - 78˚4,000 m˚< 0.05 m˚1.26 - 1.38 (Standard BK7)1.32 - 1.44 (Special N-BAF3)1.40 - 1.52 (Special NSF15)< 2 × 10 -6180 D76 Hz0.1 s - 300 s0.1 m˚Gold coated glass disk (25 mm ∅)Combined ESPR cuvetteOptional for normal SPROptional40 kg60 × 51 × 45 cm 3170 WSurface plasmon resonanceScanning mirrorCuvette system1670 nmManual and semi-automatic controlContinuous with pipette tipn.a.Syringe and peristaltic0.8 - 227 µl/s20 - 150 µl62˚ - 78˚4,000 m˚< 0.05 m˚1.26 - 1.38 (Standard BK7)1.32 - 1.44 (Special N-BAF3)1.40 - 1.52 (Special NSF15)< 2 × 10 -6180 D76 Hz0.1 s - 300 s0.1 m˚Gold coated glass disk (25 mm ∅)Combined ESPR cuvetteOptional for normal SPROptional23 kg40 × 33 × 36 cm 3170 WPAGE 7

ECO CHEMIE 09/2005KANAALWEG 29 G3526 KMUTRECHTPO BOX851633508 ADUTRECHTTHE NETHERLANDSTELEPHONE+31302893154TELEFAX+31302880715E-MAILAUTOLAB@ ECOCHEMIE.NLHOMEPAGEWWW.ECOCHEMIE.NLTHE INSTRUMENTSNEDERLAND QA B.V.THE POSSIBILITIESISO 9001: 2000THE SOFTWAREYOUR AUTOLAB DISTRIBUTOR