Centro & Centro XS³ LB 960

6 models Centro Basic upgradeable Centro Research 2 injectors (10 – 100 µl) and waste pump Centro HTS 3 injectors and waste pump, 96&384 well robot access, robot software, Mikrowin Adv. II Centro Phago 3 injectors and waste pump, 96 well, temperature control Centro XS³ Basic upgradeable Centro XS³ Reportergene 2 Injectors and waste pump, ready-to-use DLR files

Centro models & options

Features Up to 3 injectors (10 – 100 µL) Measurement of 96 & 384 well microplates Shaking Robot Accessibility Small footprint Software with calculation possibilities and GLP features, e.g. control history Heating (Centro only)

Centro XS³: Differences to Centro PMT located directly above well: better efficiency all in pre-measurement positions slightly higher housing (~10 mm) ambient temperature only higher tolerance for varying plate heights: +/- 0.6 mm Better and more stable alignment of PMT level and plate level 96 or 384 wells: change of cone

Centro XS³ optics new more direct optics

! Injectors Inj. #3 is not in measurement position any more all „slow“ flash-type assays still possible, e.g. DLR (2 sec delay between injection and reading!), single luciferase, ATP, etc. !

Performance features Sensitivity (Centro XS³) Sensitivity (Centro) ~1.1 attomol ATP (per well) Celsis SensiLux ATP kit white opaque 96 well plates Berthold ~1.3 zmol Firefly (per well) ~ 1200 molecules Promega DLR kit Sensitivity (Centro) ~4.1 attomol ATP (per well) ~1.96 zmol Firefly (per well) ~ 800 molecules

Crosstalk: < 3 x 10-5 white opaque 96 well plate Berthold < 2 x 10-6 Black opaque 96 well plate Berthold Dynamic range: > 6 decades

Method for Calculation of Detection Limits 3 x Standard Deviation (Bkg) Signal (Std) – Signal (Bkg) x Concentration (Std) Std: signal about 100 times greater than background

Detection Limit stable background is extremely important: 3 x standard deviation read a whole empty (or just reagent without analyte) plate to determine mean and SD of background read 1 sample with known concentration (or dilution series) calculate

Detection Limit

Sensitivity: Background Issues Background of an assay is usually all components of an assay; only the analyte is missing Reagents have usually a high intrinsic background; sometimes 100 – 1000 x instrument background So far all data for detection limit were derived with reagent background - the superior performance of BT instruments was compromised by those numbers Competition obviously used background values without reagent We are now in the process of reviewing / re-doing all measurements in respect of taking plain instrument background data for calculation

Sensitivity Comparison Centro - Veritas

Sensitivity calculated detection limit: Centro 312.7 pM Veritas 359.8 pM

Testplates for instruments comparison Other test plates e.g. Harta, BDS Berthold Technologies testplate Orion Veritas Centro IMPORTANT: comparison with other test plates than BT one discriminates Centro due to optical configuration

Instrument Comparisons Centro XS³ Orion Victor Lumi Star Lumino skan Lucy Veritas ATP amol Lucif. zmol 4.1 1.96 1.1 1.3 20 80 50 1 (?) 20-30 6 3 Dead vol. 800 5x600 5x1500 1200 5x1200 crosstalk 10-6 10-5 10-4 ? Waste pump Y n Inj. vol. prec. 10-100 99% 10-250 5-350 98.5% 5-250 1-1000 95% 25-500 25-250 99%? Inj. 4 2 Plate format 96 op.384 96/384 24-384 24-384* 24-384** 96 Temp. contr. N Cell inj. *insertion of spacers **manual adjustment

Centro XS³ is the direct answer to Veritas/Glomax 2 injectors direct & highly efficient optics 96 well plus: kinetics/repeated function extended calculation functions Additional injector available 384 well plate possibility

BDS Orion II Competiton PerkinElmer Victor Light Turner Veritas BMG LumiStar Labsystems Ascent Beckman Lucy III Dynex MLX

Competiton Orion II pricing: ~ 20 k€ some cases 14 k€ with 2 injectors! syringe injectors are difficult to completely empty: residues of „old“ liquids 20 amol ATP per well usually unstable background

Competiton Victor Light pricing: < 15 k€ syringe injectors in separate box dead volume 1.5 ml not usable for fast kinetics (Acridinium ester) questionable use for Aequorin assays (timing; malicious to cells) 80 amol ATP per well* high crosstalk Workout necessary for any data reduction (increased cost)

Competiton Victor Light PKI brochure: 20 pM (glow ATPLite) =20 fmol / ml =4 fmol per well (200 µl) comparison Centro: 4 amol per well !! comparison Centro XS³: exp. 1 amol per well!!! (flash Sensilux)

Competiton Turner Veritas pricing: 16 kUS$ (Jan2004) w/ 2 inj. 2 syringe injectors (min. 25 µL) to unload and clean the sytems tips have to be released and manually placed into waste container Injector tips are too close causing cross- contamination (suspending droplets) dead volume ~ 1.5 mL injectors can not be completely emptied black residues remain on plate after reading

Competiton Turner Veritas 2 dynamic range (9 decades) only via photon counting & current mode combination: takes ~ 3 sec instead of 1 sec 96 well only injector access only by opening cover currently no kinetics possible no temp control available software is primitive (DDE to EXCEL) no sophisticated calculation possible no user access (no 21 CFR part 11) runs only with US Windows OS

Competiton BMG LumiStar pricing: ~ 15 k€ 3 syringe injectors dynamic range (8 decades) with gain adjustment only Current mode of PMT lower sensitivity (~ 20 times less)

Comp. Labsystems Ascent pricing: ~ 11-20 k€ with 1 inj. & LIA software 3 syringe injectors Current mode of PMT lower sensitivity (~ 20 times less)

Comparison Asys Lucy =Beckman / =Anthos / =Mediator pricing: ??? 2 syringe injectors Current mode of PMT lower sensitivity sales through Asys will not be successful Lucy production is said to be stopped

Competiton Dynex MLX pricing: 15 – 20 k€ 3 syringe injectors: 5 µL increments only Current mode of PMT computer integrated (replacement problems after short periods of time) only 96 well plates limited sales force

Centro Selection guide

Message to take away WE ARE THE EXPERTS! We offer the best luminometers Best performance: Sensitivity Crosstalk Injector accuracy & precision Highest flexibility: Hardware Software WE ARE THE EXPERTS!