Accepted worldwide for its reliability, the FDSS is capable of 1536-well format and high-sensitivity luminescence measurements. The FDSS (Functional Drug Screening System) series are designed for cell-based assays in the drug discovery field. These instruments optically detect intracellular reactions and biological signal transmissions, and are used as screening systems to discover new lead compounds which could be candidates for new drugs. The FDSS7000EX is our high-end model capable of handling 1536-well assays and measuring both fluorescence and luminescence, and is equipped with a variety of functions such as multiple washing. Many kinds of suspended/adherent cells' real-time kinetic reactions can be measured and analyzed. Various optional parts such as FRET, robot connections, etc. are available. In addition, the FDSS7000EX is expandable for future upgrades. Applications Intracellular calcium ion Membrane potential Ion channel Aequorin Luciferase FRET Suspended cell Applications Do more with an expandable modular system Intracellular calcium ion Comparison of calcium mobilities using Aequorin, Fluo-3, and Fura-2 receptor-expressing cells. 0.7 3 5.9 pM 19.68 pM 12000 10000 8000 6000 4000 2.5 65.61 pM 0.6 218.7 pM 2 Ratio Fluorescence (RFU) Luminescence (RLU) 14000 729 pM 0.5 2.43 nM 1.5 8.1 nM 27 nM 0.4 1 90 nM 2000 300 nM 0 0.5 0 25 51 76 101 126 0.3 0 25 Time(sec) Aequorin 50 76 101 126 0 25 Time(sec) Fluo-3 50 76 101 126 1 μM Time(sec) Fura-2 Membrane potential Multiplex assays using Fura-2AM and a membrane potential fluorogenic reagent This is an example of an multiplex assay that concurrently measured membrane potential and calcium mobilization. In this example, (1) a FMP measurement with an excitation wavelength of 480 nm and a fluorescence wavelength of 540 nm and (2) a Fura-2 measurement with an excitation wavelength of 380 nm and a fluorescence wavelength of 540 nm are performed alternately, and then the mobilizations of (1) and (2) are measured concurrently. ・Fura-2AM：ex380 / em540 ・Membrane potential：ex480 / em540 Calcium and membrane potential response curves according to the changes in the ligand concentration Calcium mobilization according to the changes in the concentration of three types of antagonists with a fixed ligand concentration Ion channel Involvement of various ion channels and receptors in Ca 2+ oscillations Presented at the 8th FENS Meeting - Amsterdam - July 3-7, 2010 Session 015 Epilepsy 1, n˚015.36 2 Signals in the extremely low concentration range can also be detected by highly sensitive, luminescence sensors. ・Cell：CHO ・Substrate：h-CTZ ・Ligand：ATP ・Measurement time: The ligand is dispensed 10 seconds after measurement starts. Measured at 1 second intervals for 1 minute and 30 seconds. Control 0.3 nM 3 nM 30 nM Control 0.3 nM 3 nM 30 nM RLU (Max) Comparison of dose responses provided by an aequorin assay RLU (Max) Aequorin Time(sec) Time(sec) Fluorescence/luminescence sensor unit Fluorescence sensor unit Cell: U937 Luciferase (10 μg/mL Mitomicin-C induced apoptosis) 5000 cells/well ∼ 20 cells/well KIT: Bright-GloTM Luciferase Assay System (Promega E2610) 20 μL per well for 384 black microplate 10 min incubation at RT (25 μL per well for 384 black microplate) R KIT: Caspase-Glo Assays (Promega G8090) Add 25 μL reagent to 25 μL medium containing cells for 384 black microplate 30 min incubation at RT FRET Fluorescence resonance energy transfer (FRET) through the use of a coumarin dye (CC2-DMPE) and an oxonol dye (DISBAC). Excitation light 【FRET VSP】 Excitation light Fluorescence Fluorescence Energy transfer Vm Emission Ratio = Donor Donor Donor (CC2-DMPE) Emission (460 nm) Acceptor Acceptor (DISBAC) Emission (580 nm) Acceptor Suspended cell This is an example in which an aequorin assay was used to measure suspended cells at different CTZ concentrations. 10 000 CHO cells genetically engineered to express GPCR that can be stimulated by a neurotransmitter were dispensed to a single well. Cells loaded over 4 hours with substrate concentrations of 1 μM, 2 μM, 5 μM, and 10 μM were stimulated by ATP and ACH, which are endogenous ligands. 【ATP】 【ACH】 3 Hardware Sophisticated system structure Dispenser Can be equipped with various dispenser heads 96-, 384-, or 1536-tip dispenser heads are selectable. The tips are exchangeable. 384- and 1536-pintool heads are also supported. The main unit is equipped with an automatic tip loading feature. As standard, three types of compounds can be dispensed. If you use the back plate loading line, up to four types of compounds can be dispensed. The dispenser heads can be easily changed by the user. Pintool head Dispenser head (for tips) Pintool head (1536 and 384) NEW Dispensing involves setting metal pins in the tip positions and then attaching the reagent to the ends of the tips. This enables the dispensing of minute amounts of solution and reuse. Two dispenser heads capability Two heads can be installed at the same time, which makes it possible to use separate heads for compounds that have high viscosity or to use separate heads for agonists and antagonists. Tips When using reagents that have high viscosity Used together with pintool head Dispenser head Dispenser head 1536 head for high viscosity 1 1536 pintool head 2 for high viscosity 1536 tips Tips Tips Tips are changed each time for 1 . Normal reagent is dispensed for 2 . Washing unit Plate stacker unit Equipped with a variety of washing features Plate stacker feature for 20 or 50 plates Up to three solvents are supported, which enables washing with no carry over. Tips can be washed during measurement. An ultrasound washing feature is also available. Equipped with a wiping stage to dry tip ends. Using the plate stacker makes it possible to perform automatic measurements even without the use of a robot. You can select 20 plates or 50 plates. Stacker cassette method makes it easy to set plates. Can be equipped with a barcode reader. Wash vat Restacking function NEW OUT Wiping stage IN Restacking A stacker equipped with a feature that returns all microplates that have reached the OUT side back to the IN side is also available. Measurement Plate shaking features Others Plate shaking provides stable response detection. Auto compound feeder Measurements can be performed while applying fine vibrations to the whole plate. Cells and reagents can be mixed effectively and quickly. This is also effective for assays that use suspended cells. A new reagent feeder that drastically reduces dead volume can also be installed. Plate shaking effect for a Fluo-4 assay Temperature control function NEW The FDSS7000EX can also be equipped with a heater unit. The internal temperature sensor is used to maintain a stable temperature. Mixing OFF 4 Mixing ON Fluorescence/luminescence sensor, Fluorescence optics Supports fluorescence assays and luminescence assays with a single sensor NEW Can also be combined with a highly sensitive luminescence sensor This sensor has a fluorescence detection sensitivity that far exceeds that of conventional fluorescence sensors. Because the excitation light can be lowered, the cost of purchasing excitation light sources can be reduced. In addition to high sensitivity, the FDSS7000EX also has a high dynamic range. The fluorescence changer makes FRET measurements possible. * Conventional fluorescence sensors can also be selected. H1 NFAT HEK293 cells: Calcium measurement by Fluo-4 EC50 H1 NFAT HEK293 cells: Calcium measurement by BacMam Aequorin Histamine 1.7305e-009 Compound EC50 S/B ratio Z' Histamine 1.73 nM 19 0.66 Na-Methylhistamine 1.0661e-006 EC50 Na-Methylhistamine 1.0 μM 25 0.67 Histamine 3.8659e-009 Compound EC50 S/B ratio Z' * Reagents and data provided by Life Technologies, USA Na-Methylhistamine 3.8809e-006 Histamine 3.87 nM 142 0.84 Na-Methylhistamine 3.9 μM 182 0.68 * Reagents and data provided by Life Technologies, USA Various assays with xenon lamp & filter combination (multiple excitation and emission) Good wavelength separation by epi-illumination optics Fura-2 assay with dual excitation (340 nm & 380 nm) FRET (C/Y, Membrane potential) with emission filter changer 2 dichroic mirror and 4 emission filter slots for various assays White light xenon lamp with wattage options (150 W, 300 W, 600 W) 6 filter slots for excitation Uniform plate illumination UV Blue Fura-2(Ca2＋) SBFI(Na＋) PBFI(K＋) MQAE, DiH-MZA(Cl−) DAPI(DNA) VSP-1(FRET) Green Fluo-3 Calcium Green Sodium Green CoroNa Green BCECF GFP FITC Di-8-ANNEPS YFP JC-1 CoroNa Red Rhod-2 Rhodamine FMP Luminescence sensor Equipped with a gate function, which improves the dynamic range during luminescence measurements A highly sensitive luminescence sensor that can acquire the weak signals that cannot be acquired by the fluorescence/luminescence sensor is also available. The new gate function enables measurements of samples with a large amount of light. By combining this feature with the fluorescence/luminescence sensor, the FDSS7000EX can support an even wider range of luminescence signals. Gate function For samples that have a large amount of light, a neutral-density filter (an ND filter) is normally used. However, in this situation, only the signal is attenuated, so the signal-to-noise ratio (S/N) worsens. On the other hand, the gate function gates both the signal and the noise, so this method has a better S/N. ND filter Very weak Weak Low Luminescence sensor + Gate function Fluorescence/luminescence sensor Gate function More dynamic range Signal Noise With gate Without gate RLU NEW Luminescence signal Saturation After reduction After reduction Luminescence signal 5 Software All kinds of assays can be executed with simple operations Basic operation Select mode [Protocol mode] Set up protocol and start the assay automation. [Check mode] Check the status of the system and image. Camera/Filter control [Viewing mode] Open the measured data for analysis. Live image Set up protocol Assay protocol configuration Configure addition, wash frequency and sequence in measurement Automation from measurement to data output Dispensing protocol Configure height, dispense speed, and tip mixing Plate shaking On/Off Get results Measurement / Analysis Displays the kinetic data, auto-scale graph, correlation diagram, and analysis results for all wells Equipped with a variety of analysis features and output features Data analysis Data output • Negative control • Raw data • Positive control • Text • Subtract bias • Bitmap (graph) • Spatial uniformity • Peak calculation • Hit identification • IC/EC calculation ▲ Measured data 6 Detailed settings Assay protocol configuration Up to 4000 samples can be taken for one protocol. Alternatively switch between two sampling intervals (up to three times). Graph Correction Range Analysis Use the tabs to configure a variety of settings. Dispensing protocol Settings such as the dispensing amount and speed can be configured. Settings can also be configured for operations such as washing and tip trituration. Up to four additions can be performed in a single protocol! addition 4 addition 3 addition 2 addition 1 7 System configuration Flexible system for diverse needs Fluorescence measurements Calcium ion assays are in ever increasing demand in areas such as lead discovery, pharmacological study, and food product function research. The FDSS7000EX supports various types of ion channel assays such as Na+, K+, Cl-, and pH as well as calcium ion assays using fluorescent dyes such as Fura2, Fluo-3, and Fluo-4. FDSS7000EX main unit Washing unit Ligand plate Dispenser head Barcode reader OUT Microplate stacker Excitation light source unit Fluorescence sensor unit IN Microplate stacker front Dispenser tip Fluorescence/Luminescence The fluorescence/luminescence sensor is capable of detecting both types of emission. Adding a filter changer enables FRET measurements. FDSS7000EX main unit Washing unit Ligand plate Dispenser head Barcode reader OUT Microplate stacker Excitation light source unit Fluorescence/luminescence sensor unit IN Microplate stacker front Dispenser tip * For weak-luminescence detection, use an ultrasensitive luminescence sensor with even higher sensitivity. FRET measurement The FDSS7000EX is utilized in assays where two different fluorescence wavelengths are measured, for example FRET probes (VSP-1, Premo TM), cAMP dyes (FICRhR), simultaneous calcium and membrane potential dyes (multiplexing), and dual reporter gene assays (GFP and DsRed). FDSS7000EX main unit Washing unit Ligand plate Dispenser head Barcode reader OUT Microplate stacker Excitation light source unit Fluorescence/luminescence sensor unit IN (or Fluorescence sensor unit) Microplate stacker front FRET optics exchanging device Dispenser tip FRET optics exchanging device A8472-04 The A8472-04 automatically switches between the fluorescence filters and the dichroic mirrors. This device is used for assays that handle dual-wavelength fluorescence such as FRET or VSP. Filter switching interval * 0.3 s Number of installed dichroic mirrors Max. 2 Number of installed fluorescence filters Max. 4 * Switching interval between adjacent filter positions 8 FDSS7000EX main unit DM1 DM2 FDSS7000EX main unit DM1 DM2 Changes the acquired wavelength by rotating the fluorescence filter Dichroic mirror Sensor Fluorescence filters Controller Sensor Controller Fluorescence and luminescence measurement The standard configuration has three compound/ligand plate positions. By adding the rear loader, you can have the fourth compound/ligand plate for dispensing. The rear loader plate can be kept inside the FDSS7000EX and can be used for the next protocol if required. (4-reagent dispensing structure) FDSS7000EX main unit Barcode Microplate reader stacker Washing unit OUT IN Microplate stacker rear Ligand plate Dispenser head Barcode Microplate reader stacker OUT IN Microplate stacker front Excitation light source unit Fluorescence/luminescence sensor unit (or Fluorescence sensor unit) FRET optics exchanging device Dispenser tip Luminescence measurement The FDSS7000EX is ideal for calcium ion assays using a calcium-sensitive photoprotein "aequorin" along with reporter gene assays that make use of luciferin-luciferase reactions, high-sensitivity protein interaction analysis (two-hybrid assays), and luminescence experiments where the amount of light emissions cannot be predicted. FDSS7000EX main unit Barcode Microplate reader stacker Washing unit OUT IN Microplate stacker rear Luminescence sensor unit Ligand plate Dispenser head Dispenser tip Fluorescence and luminescence measurements The FDSS7000EX is a stand-alone system integrated with powerful functions for cell-based kinetic assays, and capable of detecting a wide variety of fluorescent and luminescent phenomena utilizing 96, 384, and 1536 well plates. FDSS7000EX main unit Barcode Microplate reader stacker Washing unit OUT IN Microplate stacker rear Luminescence sensor unit Ligand plate Dispenser head Barcode Microplate reader stacker OUT IN Microplate stacker front Excitation light source unit Fluorescence/luminescence sensor unit (or Fluorescence sensor unit) FRET optics exchanging device Dispenser tip Robot connection Interface for connecting a robot is available. Microplate 1 2 3 Plate from the robot system to FDSS plate loading arm FDSS measurement by robot control After the measurement, robot takes out plate from the FDSS plate unloading arm Manufacturers of compatible products Thermo Scientific, Beckman Coulter, HighResBiosolution, Hamilton, Agilent FDSS7000EX main unit 1 Robot system 3 Plate loading arm Data analysis system 2 - Change the assay protocol - Start measurement Control signal Serial I/F 9 System configuration A rich variety of optional parts makes it possible to design a system that meets your specific needs Component Dispensing unit One-unit and two-unit versions are available Equipped with an automatic tip loading feature Pintool head Washing unit A one-solvent washing version (equipped with an ultrasonic washing feature) to a three-solvent washing version are available Equipped with an automatic wiping stage Microplate stacker Fluorescence sensor unit Fluorescence/luminescence sensor unit *1 96 tip type dispensing unit (10-200 μL) 384 tip type dispensing unit (1-30 μL) 1536 tip type dispensing unit (1-5 μL) 384 pintool head for short pins 384 pintool head for long pins 1536 pintool head for short pins 1536 pintool head for long pins Washing unit Additional washing unit 96 chimney plate 384 chimney plate 1536 chimney plate Microplate stacker front (without lid opener) Microplate stacker rear (without lid opener) Microplate stacker front (lid opener) Microplate stacker rear (lid opener) Microplate restacker front (without lid opener) Microplate restacker rear (without lid opener) Single plate holder Microplate stacker cassettes Microplate stacker cassettes Fluorescence sensor unit (1 lamp) Fluorescence sensor unit (2 lamps) Fluorescence sensor unit (4 lamps) Fluorescence/luminescence sensor unit (1 lamp) Fluorescence/luminescence sensor unit (2 lamps) Fluorescence/luminescence sensor unit (4 lamps) Luminescence sensor unit for 1536 wells FRET optics exchanging device Barcode reader Auto cell feeder Heater unit *1 *2 *1 To use the fluorescence/luminescence sensor and the heater unit at the same time, the FDSS7000EX requires a water cooling device. For details, contact your Hamamatsu sales representative. *2 You can only select the heater unit at the time of purchase. Consumable Tips Tips for 96 wells (10 racks) Tips for 384 wells (10 racks) Tips for 1536 wells (10 racks) A8687-32 A8687-62 A8687-52 Optical filter * See the back page for details. Maintenance To maintain long-term stability of this device, we recommend that you purchase the maintenance contract. For details, contact your Hamamatsu sales representative. 10 A10118-24 A10118-26 A10118-28 A10118-30 A10118-31 A10118-27 A10118-29 A10118-47 A10118-48 A10118-44 A10118-43 A10118-46 A10118-04 A10118-05 A10118-94 A10118-95 A10118-07 A10118-08 A10118-96 A10118-92 A10118-93 C10512-11 C10512-12 C10512-13 C11653-11 C11653-12 C11653-13 C10520-11 A8472-04 A10118-50 A10118-55 A10118-09 Dispensing unit 1536 format Dispensing unit A10118-28 Tips A8687-52 Chimney plate A10118-46 Tips A8687-62 Chimney plate A10118-43 Tips A8687-32 Chimney plate A10118-44 384 format Dispensing unit A10118-26 96 format Dispensing unit A10118-24 Pintool head (for 384/1536) * In this photograph, pins are already attached. Note that the pins are not included. Pump unit Excitation light source unit Excitation light source unit (150 W) Excitation light source unit (300 W) Pump unit Filter changer Microplate stacker Microplate stacker Excitation light source unit (600 W) Microplate stacker cassettes Filter changer 11 System configuration A rich variety of optional parts makes it possible to design a system that meets your specific needs Consumable Optical filter Standard Option Application Excitation filter 1 Excitation filter 2 Dichroic mirror Emission filter 1 Emission filter 2 Fura-2 (Ca2+) 340 nm 387 nm UV 520 nm to 560 nm - SBFI (Na+) 340 nm 387 nm UV 520 nm to 560 nm - PBFI (K+) 340 nm 387 nm UV 520 nm to 560 nm - MQAE, diH-MQA (Cl-) 360 nm - UV 440 nm to 470 nm - DAPI (DNA) 360 nm - UV 440 nm to 470 nm - VSP-1 387 nm - for VSP-1 465 nm 565 nm CFP/YFP 438 nm - for C/Y 483 nm 542 nm Fluo-3, Fluo-4, Fluo-8 (Ca2+) 472 nm - B 520 nm to 560 nm - Calcium Green (Ca2+) 472 nm - B 520 nm to 560 nm - Sodium Green, CoroNa Green (Na+) 472 nm - B 520 nm to 560 nm - BCECF (pH) 472 nm 450 nm B 520 nm to 560 nm - GFP 472 nm - B 520 nm to 560 nm - FITC 472 nm - B 520 nm to 560 nm - YFP 500 nm - for Y 542 nm - Di-8-ANNEPS 472 nm - B 645 nm long pass - JC-1 540 nm - for JC-1 570 nm to 600 nm - CoroNa Red (Na+) 531 nm - for FMP 560 nm to 640 nm - Rhod-2, Rhod-3, Rhod-4 (Ca2+) 560 nm - for FMP 590 nm to 650 nm - Rhodamine 560 nm - for FMP 590 nm to 650 nm - FMP 531 nm - for FMP 593 nm - * Above wavelength are not representing the filter characteristic, just showing the general wavelength for each application. The filters we provide are the best-optimized selection and combination with our optics, and does not match the wavelength shown here. Dimensions/Weight Approx. 1500 mm (W) × 1030 mm (D) × 1450 mm (H) Main unit Up to approx. 300 kg 600 mm (W) × 700 mm (D) × 1500 mm (H) Data analysis system Approx. 50 kg 500 mm (W) × 500 mm (D) × 1200 mm (H) Pump unit / Tank Approx. 50 kg System footprint Pump unit / Tank Main unit Excitation light source Data analysis system unit 1300 mm Microplate stacker 2800 mm FDSS is registered trademark of Hamamatsu Photonics K.K. Product and software package names noted in this documentation are trademarks or registered trademarks of their respective manufacturers. Subject to local technical requirements and regulations, availability of products included in this promotional material may vary. Please consult with your local sales representative. Information furnished by HAMAMATSU is believed to be reliable. However, no responsibility is assumed for possible inaccuracies or omissions. Specifications and external appearance are subject to change without notice. © 2014 Hamamatsu Photonics K.K. HAMAMATSU PHOTONICS K.K. www.hamamatsu.com HAMAMATSU PHOTONICS K.K., Systems Division 812 Joko-cho, Higashi-ku, Hamamatsu City, 431-3196, Japan, Telephone: (81)53-431-0124, Fax: (81)53-435-1574, E-mail: [email protected] U.S.A.: Hamamatsu Corporation: 360 Foothill Road, Bridgewater, N.J 08807, U.S.A., Telephone: (1)908-231-0960, Fax: (1)908-231-1218 E-mail: [email protected] Germany: Hamamatsu Photonics Deutschland GmbH.: Arzbergerstr. 10, D-82211 Herrsching am Ammersee, Germany, Telephone: (49)8152-375-0, Fax: (49)8152-265-8 E-mail: [email protected] France: Hamamatsu Photonics France S.A.R.L.: 19, Rue du Saule Trapu, Parc du Moulin de Massy, 91882 Massy Cedex, France, Telephone: (33)1 69 53 71 00, Fax: (33)1 69 53 71 10 E-mail: [email protected] United Kingdom: Hamamatsu Photonics UK Limited: 2 Howard Court,10 Tewin Road, Welwyn Garden City, Hertfordshire AL7 1BW, UK, Telephone: (44)1707-294888, Fax: (44)1707-325777 E-mail: [email protected] North Europe: Hamamatsu Photonics Norden AB: Torshamnsgatan 35 16440 Kista, Sweden, Telephone: (46)8-509-031-00, Fax: (46)8-509-031-01 E-mail: [email protected] Italy: Hamamatsu Photonics Italia S.r.l.: Strada della Moia, 1 int. 6 20020 Arese (Milano), Italy, Telephone: (39)02-93581733, Fax: (39)02-93581741 E-mail: [email protected] China: Hamamatsu Photonics (China) Co., Ltd.: B1201 Jiaming Center, No.27 Dongsanhuan Beilu, Chaoyang District, Beijing 100020, China, Telephone: (86)10-6586-6006, Fax: (86)10-6586-2866 E-mail: [email protected] Cat. No. SBIS0050E07 DEC/2014 HPK Created in Japan