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Tecan’s monochromator innovations for microplate reading Tecan’s Quad4 Monochromators™ – Stray light reduction improves sensitivity

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  • Tecans monochromator innovations for microplate readingTecans Quad4 Monochromators Stray light reduction improves sensitivity

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    Tecans Quad4 Monochromators Stray light reduction improves sensitivity

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    Tecan DetectionTecan has a solid reputation in producing innovative and high quality automated liquid handling workstations and detection instruments such as microplate readers, microarray related products and washers. Tecan has more than two decades experience in the design, development and production of microplate readers, won several innovation prices and is ISO 9001 and ISO 13485 certified.

    Tecan has developed, produced and sold more than 20,000microplate readers globally to academic centers, governmentalinstitutions, universities, the life science, biotech andpharmaceutical industries and the Companys OEM partners. The robustness and longevity of older generation instruments

    including the Sunrise absorbance reader, the Ultra, the GENios Pro and the Safire2 is such that many have been, and still are, in daily use in biopharmaceutical screening and research facilities around the world.

    The Company has introduced a broad range of technologyinnovations such as fluorescence lifetime detection andQuad4 Monochromators. For Tecan Austria, a very important focus has been the development of the Infinite series of multimode microplate readers and, in particular, monochromator-based multimode microplate readers. These innovations have helped shape the life sciences industry, and will continue to do so.

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    Monochromator Technologies and products

    What is a monochromator?

    A monochromator is an optical instrument consisting of anentrance slit, a dispersive element and an exit slit. The dispersiveelement diffracts the light into its optical spectrum, and projects it onto the exit slit.

    Rotating the dispersive element moves the spectrum acrossthe exit slit to select the desired wavelength. In principle, thedispersive element could be realized by using glass or silica prisms,however, Tecans monochromators are designed with state of theart concave holographic gratings for highest efficiency.

    What is the advantage of a double monochromator?

    A single monochromator achieves a stray light suppression of approximately 103. If a monochromator is set for light with awavelength of 500 nm and the detector detects a signal of 10,000counts, light with wavelengths other than 500 nm may create asignal of up to 10 counts. For fluorescence applications, stray lightsuppression of single monochromators might be compromising,as the fluorescent light to be detected is usually much weakerthan 1/1000 of the excitation light.

    To achieve a higher level of blocking, Tecan has connected twomonochromators in series. Therefore, the exit slit of the firstmonochromator simultaneously acts as the entrance slit of thesecond monochromator. Size and placement of the slits in thelight path determine the effective wavelength resolution. Bycombining two monochromators, the stray light suppressionreaches a factor of 106.

    A stream of innovation for your success

    The picture above shows the history of Tecans monochromator-basedmultimode microplate readers. The development of Tecans first Quad4Monochromators-based multimode reader the Safire* startedin 1999, and the Infinite M200 PRO and Infinite M1000 PRO are Tecanscurrent monochromator-based products.

    *The Safire has now been phased out.

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    The Quad4 Monochromators technologymakes use of double monochromators forboth excitation and emission. The picturesoutline the double monochromatorsystem architecture on the excitation (left)and the emission sides (right). A sphericalmirror maps the light onto the entry slit,and the grating causes diffraction of thelight into its optical spectrum. Rotation ofthe grating determines which color willpass through the exit slit, which issimultaneously the entry slit for thesecond monochromator. The light is thenfurther diffracted by the second grating.

    What is the difference between additive and subtractive monochromators?

    In an additive monochromator design, combining two monochromators also doubles the dispersion of the system. For a given slit width, only half of the bandwidth and half of the energy is able to pass through the system. To increase monochromator efficiency, Tecans current monochromators use a special folding of the optical axis to achieve a subtractive monochromator design, causing dispersion of the first monochromator to be canceled out by the second monochromator. A subtractive monochromator design behaves similarly to tunable filters, achieving very good stray light suppression, without the transmission loss of additive monochromators.

    Tecans Quad4 Monochromators technology

    Tecan started development of its first monochromator-basedmultimode microplate reader in 1999, resulting in the launchof the Safire microplate reader. This was Tecans firstmonochromator-based multimode microplate reader tofeature the Quad4 Monochromators design.This concept places two monochromators (spectral gratings)in series on the excitation as well as on the emission side.

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    Tecans premium Quad4Monochromators technology

    Tecan has further optimized its subtractive Quad4 Monochromatorstechnology to provide more light to the sample and the detector.Premium Quad4 Monochromators feature an improved spectralblocking and stray light reduction down to 2x10-7. Used in theInfinite M1000, this is combined with a flash lamp providing 2xmore optical power, and a switchable flash frequency (patent US 7,088,054) function to optimize TR-FRET based measurements.

    In addition to Quad4 Monochromators technology, there is anenormous amount of experience and know-how which flows into thedesign integration and fine-tuning of Tecans monochromator-basedmultimode microplate readers. Tecan uses the subtractiveQuad4 Monochromator principle in both its Infinite M200 PRO andInfinite M1000 PRO multimode readers, providing monochromator-based instruments to suit a variety of market segments.

    Tecans Quad4 Monochromatorsmultimode microplate reader chronology

    Tecan has over a decade of experience in development ofmonochromator-based microplate readers and related innovations.

    Tecan started the development of its first Quad4 Monochromator-based multimode microplate reader in 1999. The Safire multimodereader was launched in 2001, offering absorbance scans,fluorescence excitation scans (emission fixed, excitation range),fluorescence emission scans (excitation fixed, emission range),3D scans (excitation and emission range), wavelength selection in1nm steps, z-focus, optional barcode reading, 1536-well reading and adiode array detector option for high speed absorbance scanning. TheSafire was Tecans first multimode microplate reader with an additiveQuad4 Monochromators design, and featured four adjustable slitssettings (2.5, 5.0, 7.5, 12.0 nm) for fluorescence-based measurements.Adjustable bandwidth settings optimize the performance of theinstrument for resolution and sensitivity by controlling the amount of light in fluorescence mode.

    Monochromator innovation

    The Quad4 Monochromators technology provides a number ofessential benefits by combining flexibility with sensitivity throughoptimized stray light reduction. The monochromator-based Infinitemicroplate readers do not require any absorbance or fluorescencefilters, offering freedom of wavelength selection, convenience andproviding a cost-effective solution when a broad variety offrequently changing applications are covered. True wavelength flexibility Using Quad4 Monochromators enables good performance

    over the whole wavelength spectrum.

    Improved sensitivity Stray light and the resulting background noise are the

    most critical factors affecting sensitivity. Tecans Quad4 Monochromators effectively reduces stray light down to 10-6.

    More accurate readings Using Quad4 Monochromators gives a true spectral scan,

    by eliminating the use of cut-off filters.

    Extremely fast scan times Using Tecans configuration of Quad4 Monochromators

    increases the speed of fluorescence scans compared with microplate readers combining two monochromators with cut-off filters.

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    With the Safire2 instrument, Tecan introduced the subtractive Quad4 Monochromators technology. The Safire2 offered the option of multichannel absorbance and luminescence reading, as well as fluorescence polarization and an integrated stacker. The number of adjustable bandwidth settings was also extended, to 16 slit options on each the excitation and emission side, providing an enormous range of potential wavelength/bandwidth settings for assay development. Tecan also developed the patented stepless adjustable bandwidth setting (patent US 7,123,398).

    With the Infinite M1000, Tecan launched the premium Quad4 Mononchromators, which features a subtractive monochromator design, an integrated solution to provide more light to the sample and detector and improved stray light reduction down to 2x 10-7.

    The Infinite M200 offered affordable access to subtractive Quad4 Monochromators technology. The Infinite M200 was relaunched in 2009, with enhanced monochromator performance (accuracy) for DNA quantitation (260/280 nm) and absorbance scanning. The Infinite M200 was also compatible with the recently launched NanoQuant Plate, which allows DNA quantitation in low sample volumes down to 2 l.

    The new Infinite M200 PRO provides advanced FI bottom reading with optimal read (OR) function and is designed to optimize cell-based measurements. Very low CVs and high intra-/inter-well reproducibility can be achieved when measuring adherent cells in microplates, offering increased sensitivity. Automated, adjustable z-focus facilitates assay miniaturization for FI Top measurements , achieving equally high sensitivity and performance in low volume plates. In addition, advanced background correction makes the instrument particularly suited to cell-based applications using autofluorescent growth media, providing automatic optimization of the signal-to-noise ratio.

    The new Infinite M1000 PRO is available with a module designedspecifically for AlphaScreen and AlphaLISA technologies, and equipped with a high power laser light source. The AlphaScreen module includes dedicated filters for AlphaScreen and AlphaLISA assays, as well as an innovative temperature correction function that compensates for sample temperature variations across the microplate. Additionally, the Infinite M1000 PRO also offers luminescence scanning to complement its existing 3D and ultra fast absorbance and fluorescence scanning capabilities.

    A decade of monochromator-based innovations in multimode microplate reading

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    wavelength range. The new Infinite M1000 PRO enables fast absorbance scans (multi-channel capability), fluorescence scans(excitation, emission) and 3D scans (allowing serial emission scans at different excitation wavelengths). This powerful tool helps to identify changes in the spectral properties of fluorescent probes or the characterization of unknown fluorescent sample. A new feature is luminescence scanning, recording the emission spectra of stable luminescent signals at 1 nm resolution using the instruments emission monochromators.

    Outstanding flexibility, sensitivity and speedThe modular Infinite M1000 PRO can be used with 6- to 1,536-wellmicroplates, cuvettes and Tecans NanoQuant Plate, and isequipped with Instant Read controls for rapid start/stop of preprogrammed measurements. Its Optimal Read function for FI bottom reading offers enhanced performance for cell-basedapplications, performing multiple measurements on spatiallyseparated spots arrayed across each well area, achievingmaximum illumination and sensitivity. The Infinite M1000 PROalso features AlphaScreen/AlphaLISA measurements, luminescence color-filters and automated z-focusing foroptimization of signal-to-blank ratios in all top reading modes.

    The Infinite M1000 PRO is upgradable, can be equipped with abarcode scanner, reagent-saving injectors, an integrated stacker option and is able to be fully integrated into Tecans Freedom EVO liquid handling workstations.

    Empowerment for advanced life sciences and drug discovery

    Premium Quad4 MonochromatorsThe Infinite M1000 PRO plate reader features premium Quad4 Monochromators technology. Using a flash lamp with double the power and switchable flash frequency settings for fluorescence intensity, it provides even more light to both the sample and the detector, as well as offering improved spectral blocking and extended stray light reduction down to 2x10-7.

    The Infinite M1000 PRO also features adjustable bandwidths for fluorescence-based measurements: 16 possibilities for each excitation and emission wavelength selection, providing a total of 256 potential fixed excitation/emission wavelength pairs. Adjustable bandwith can be applied in fluorescence intensity (FI), fluorescence polarization emission (FP; only adjustable bandwidth on emission side), fluorescence resonance energy transfer (FRET), time-resolved fluorescence (TRF), time-resolved fluorescence resonance energy transfer (TR-FRET) and fluorescence scanning modes. Optimizing monochromator bandwidth results in improved instrument sensitivity, particularly for dyes with a small Stokes shift and advanced multiplexed testing.

    Advanced scanning capabilitiesThe Infinite M1000s premium Quad4 Monochromators offer theflexibility to scan novel or unknown probes across a broad

    NEW Infinite M1000 PRO

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    Empowerment for basic research and life sciences

    Tecans many years of experience in monochromator design haslead to a surprisingly powerful yet affordable multimode reader,making this sophisticated technology accessible for basic researchand life science applications.

    The easy to use and upgradable Infinite M200 PRO featuressubtractive Quad4 Monochromators, offering a choice ofdifferent detection modes like eg absorbance and fluorescenceintensity (both UVNIR and wavelength scanning), TRF andluminescence (glow/flash) measurements.

    Sensitivity and format flexibility The instrument provides a good solution for a broad range ofapplications requiring excellent sensitivity, multiplexing capabilityand high format flexibility, including 6- to 384-well microplates,and cuvettes.

    Since its introduction in 2006, the Infinite series has beenoptimized for its monochromator speed and accuracy, and isideally suited to a broad range of applications. It was the firstmultimode reader capable of absorbance measurements with lowsample volumes, and is compatible with Tecans patented (EP2045015) NanoQuant Plate, an innovative quartz-based tool that can measure up to 16 samples simultaneously from just 2 l sample volumes.

    Infinite M200 PRO

    The combination of this sensitive plate reader and theNanoQuant Plate makes the Infinite M200 PRO one of the mostaccurate and reproducible multimode monochromatorinstruments on the market.

    Dedicated cell-based reader Features such as optimal read (OR) and automated z-foucs withadvanced background correction make the Infinite M200 PROideal for cell-based applications, particular when usingautofluorescent growth media. In addition, the highly innovativeGas Control Module (GCM) offers external control of CO2 and O2,helping to maintain stable culture conditions during measurements and allowing assays to be performed underanaerobic or physiological conditions. This extends the experimental window while ensuring consistent and biologicallyrelevant data.

    The Infinite M200 PRO can also be equipped with an injector,can be used with the Connect stacker system for batch processingof up to 50 plates, or can be integrated into Freedom EVO liquidhandling workstations.

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    The current Infinite M1000 PRO model with premium Quad4Monochromators builds on these innovative technologies,supplying more light to the sample and the detector to allow theuse of advanced assay formats in drug discovery, life sciences andresearch.

    Tecan also provides access to affordable Quad4 Monochromatorstechnology in form of the Infinite M200 PRO model, whichfeatures a subtractive Quad4 Monochromators design offeringimproved performance in absorbance mode for DNA quantitation(260/280) endpoint measurements and absorbance scanning.

    So whatever your monochromator-based multimode readerneeds, Talk to Tecan to find the right reader for your applicationand budget, relying on more than 10 years of know-how, competence and innovation to boost your research.

    Tecan is a well known provider of detection instrumentation,particularly multimode microplate readers, and has developed astream of innovative products and technologies to create a strongbrand and good reputation in this sector.

    Monochromator-based multimode microplate reader technologycontinues to be a key focus area for Tecan, building on a decadelong trend of successful development in the field.

    Tecan first launched the powerful Quad4 Monochromatorsconcept in 2001 with the Safire, initially using an additivemonochromator design and 4 adjustable bandwidth settings.With the Safire2 model, the number of bandwidth settings wasextended to 16, and Tecan changed to a subtractive Quad4Monochromators design.

    Customer perspective

    Tecans monochromator microplate reader portfolio. For more information www.tecan.com/Quad4

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    Flexibility, convenience and performance for your multimode microplate reading

    Tecan, Infinite and Freedom EVO are registered trademarks and Quad4 Monochromators, NanoQuant Plate, Instant Read, Sunrise, Ultra, GeniosPro, Safire and Connect are trademarks of Tecan Group Ltd. Mnnedorf, Switzerland.AlphaScreen and AlphaLISA are registered trademarks by PerkinElmer, USA. HTRF is a registered trademark of Cisbio bioassays, France. The fluorescence ratio associated with the HTRF readout is a correction method developed by Cisbio international and covered by the US patent 5,527,684 and its foreign equivalents, for which Cisbio international has granted a license to Tecan Group Ltd. Its application is strictly limited to the use of HTRF reagents and technology, excluding any other TR-FRET technologies. Transcreener HTS Assay Platform is a patented technology of BellBrook Labs, LLC USA. Transcreener is a registered trademark of BellBrook Labs, LLC USA.DLReady is a trademark of Promega Corporation, USA. LanthaScreen is a registered trademark of Invitrogen (life technologies) Corporation, Carlsbad, USA.

    2012, Tecan Trading AG, Switzerland, all rights reserved. For disclaimer and trademarks please visit www.tecan.com

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