Productivity Barriers

Transcription

1 LumiLux Cellular Screening Platform Break Through the Productivity Barriers in Cellular Screening

2 Imagine an instrument that will cellular screening productivity Introducing a high impact instrument whose technology breaks through the time, cost and performance barriers that have prevented meaningful productivity gains in cellular screening. The new LumiLux Cellular Screening Platform with its innovative cell stirrer enables luminescent cellular screening with both suspension and adherent cells, providing an ultra high throughput platform that delivers faster, superior screens right away. Key Applications Flash luminescence assays Functional GPCR calcium assays Calcium ion channel assays Tyrosine kinase receptors Glow luminescence assays Reporter gene assays The LumiLux is an ultra high throughput cellular imaging platform for both kinetic flash and glow luminescence assays with advanced liquid handling functionality. An instrument with walk-away capability, it provides sensitive detection based on patented fiber-optic imaging technology, state-of-theart liquid handling and sophisticated data analysis tools all integrated into one advanced solution for cellular assays. Plus, LumiLux introduces a breakthrough advance a unique cell stirrer that permits the use of suspension cells in uhts screens. With LumiLux you ll generate superior quality screening data (low backgrounds, higher Z's and no interference from inherent compound fluorescence) using fast, cost-effective assay methodologies. Particularly suited for uhts calcium screening, you can now combine aequorin technology with the simplicity of suspension cell assays in a 1536-well format, allowing your screening to reach unprecedented levels of productivity. This is the high impact you ll get with the robust LumiLux Cellular Screening Platform. PerkinElmer wishes to thank Euroscreen for providing a CHO-H1 cell line for use in aequorin assays. 2

3 solve your challenges virtually overnight! The LumiLux Difference Switch your calcium fluorescence assays to luminescence and significantly reduce your labor and consumable costs. At the same time, markedly increase your productivity and screening throughput. With LumiLux, you can miniaturize to 1536-well format and run continuously 5 days a week! How LumiLux is Different Luminescence Detection Technology Performs both flash and glow luminescence assays. Works with all aequorin assay methodologies (suspension cells and adherent cells). Interference from autofluorescence compounds is greatly reduced. Works with fluorescent compounds (e.g., search for an antagonist when the agonist is fluorescent). Works with suspension cells as well as adherent cells Runs flash luminescence assays, e.g. aequorin, using suspension cells in up to 1536-well formats using our unique cell stirrer. Novel cell stirrer keeps large quantities of cells in suspension. How It Helps You Reduces repeat testing from false positives. Reduces reagent and labware costs using aequorin luminescence technology vs. fluorescence technology. Improves the quality of your screening results. Requires fewer cells, reagents, labware and less labor. You realize immediate savings in screening costs. Prepare only one batch of cells/day. Eliminates the time required to grow adherent cells in microplates prior to an assay. Lets you run assays 5 days a week with no weekend work, increasing your productivity. Microplate flexibility up to 1536 Handles 96-, 384- and 1536-well microplates. Requires no hardware changes to miniaturize your assays. Switches between assay formats without changes in instrument setup. Maximum flexibility and instrument utility greatly increases throughput and productivity with lower costs. Enables assay development and screening on the same platform. Fully automated, walk away platform Runs all assay steps compound addition, cell addition, agonist/ antagonist additions, plate lid removal, timed incubations, etc. Integrated proven Packard liquid handling functionality enables complete processing of samples from serial dilutions in 96- and 384-well plates to compound preparation prior to a screen. Lets you run assay with no manual intervention just load your compound plates and cells, and go! Completely automates labor-intensive and repetitive tasks, adding key functionality. Two instruments in one Detector and Liquid Handler! Proprietary, highly sensitive camera Patented fiber-optic contact imaging provides 4.5 logs dynamic range, highly efficient light gathering and low crosstalk (< 1% for 1536). There are no parallax effects, no problems with reflection and no focusing requirements. Capable of working in the red and blue spectral regions. Sensitive detection reduces the number of cells and substrate required, reducing costs and labor. Spectral range offers application flexibility. continued on page 4 3

4 How LumiLux is Different (continued) Multi-tasking capability Runs multiple tasks at the same time, with true parallel processing capabilities. While measuring a plate, other cell or compound plates can be moving along the conveyor and onto the deck, tips can be washed or compounds/cells aspirated ready for the next assay addition. How It Helps You (continued) Speeds up sample processing and maximizes throughput. Integrated data handling and SQL server database AssayPro Includes ready-to-go protocols to get you started. Organizes, stores and analyzes data. Runtime and off-line data analysis tools. Analyzes your data in real time to identify hits and track Z's. Lets you work offline to calculate concentration response curves and EC 50 s. Reduce the Time and Cost of Calcium Screening with LumiLux Plus Aequorin Fluorescence Calcium Screening Protocol Luminescence Aequorin Calcium Screening Protocol DAY 1 Harvest cells DAY 1 Harvest cells Seed cells into tissue culture treated microplates Load cells with Coelenterazine DAY 2 Aspirate media and load cells with fluorescent dye Wash (optional dependent on assay kit being used) Dilute cells. Inject into compound plates and read or inject compounds, then cells, and read Inject compounds and read Luminescence aequorin calcium screening with LumiLux is faster than fluorescence methods (one day versus two), less costly and higher throughput. Assays are more robust and data quality is improved, proven by higher Z's and fewer false positives from autofluorescent compounds. 4

5 Faster, superior calcium screening with luminescence aequorin assays on LumiLux LumiLux provides a simpler, more robust and higher quality alternative to other calcium screening platforms, making it possible to maximize the potential of aequorin technology for the first time. It delivers higher sensitivity and lower background assays, resulting in higher Z's. And because LumiLux lets you work with suspension cells, you can reduce cell culture costs, labor and equipment use; simplify your assay protocol; and achieve even higher resource utilization and throughput every day of the week. LumiLux significantly increases calcium assay throughput using 384 inject and read capability, integrated stackers, cell stirrer and multiple deck positions to automate up to 150 x 384-well plates and 200 x 1536-well plates screening for agonist or dual agonist/antagonist assays without manual intervention. Plus only LumiLux lets you miniaturize luminescence cellular assays to 1536 wells increasing capacity and throughput even more. It all comes down to better, faster calcium screening. CHO-H1 Aequorin Assays Using Suspension Cells Using suspension cells and aequorin technology, you can markedly improve the performance and throughput of your 384-well calcium screens compared to fluorescent assays with adherent cells. When you are ready to miniaturize to 1536 wells, LumiLux can take you there. Because of the sensitivity of the LumiLux platform, assays can be miniatured to 1536-well format to increase throughput with no impact on the data being generated. This also further reduces cell culture costs since so few cells are used in these assays. 384-well Aequorin Data Using CHO-H1 Cell Line 384-well Agonist Dose Response Curves 384-well Antagonist Dose Response Curve Concentration response curves for Histamine and ATP (384-well suspension assay, 12,500 cells per well). Data from a representative experiment are shown as mean +/- SD from 8 wells. Inhibition curve for the antagonist trans-triprolidine (384-well suspension assay, 12,500 cells per well). Data from a representative experiment are shown as mean +/- SD from 16 wells. Control Z' pec 50 /pic 50 CRC Z' Histamine ATP trans-triprolidine Z' = 1- (3 x SD N + 3 x SD P) / (Ave P - Ave N) continued on page 6 5

6 384-well Aequorin Data Using CHO-H1 Cell Line (continued from page 5) Effect of cell number on signal and Z' for aequorin assay using Histamine-stimulated CHO aequorin H1 cells in 384-well suspension assay Data from a representative experiment are shown as mean +/- SD from 8 wells. The data demonstrates that the number of cells can be significantly reduced with no impact on the quality of the assay well Aequorin Data Using CHO-H1 Cell Line Kinetic Data Display Effect of cell number on signal and Z' for aequorin assay using Histamine-stimulated CHO aequorin H1 cells in 1536-well suspension assay Cells/Well Plate Z' S/B* pec 50 CRC Z' 1, , Kinetic data display for an aequorin assay using CHO-H1 cells in 1536-well format. The data demonstrates that the number of cells can be significantly reduced with no impact on the quality of the assay. Data from a representative experiment are shown as mean +/- SD from 16 wells. *S/B = Signal (Average of positive control)/background (Average of negative control) 1536-well Agonist Dose Response Curves 1536-well Antagonist Dose Response Concentration response curves for Histamine and ATP (1536-well suspension assay, 1,500 cells/well). Data from a representative experiment are shown as mean +/- SD from 32 wells. Inhibition curve for antagonist trans-triprolidine (1536-well suspension assay, 1,500 cells/well). Data from a representative experiment are shown as mean +/- SD from 64 wells. Control Z' pec 50 /pic 50 CRC Z' Histamine ATP trans-triprolidine

7 Aequorin Assay with Transiently Transfected Cell Line LumiLux can also be used to detect and screen aequorin responses in transiently transfected cell lines, demonstrating its robustness and sensitivity in measuring low responding cells. With LumiLux, you can readily use aequorin technology in assay development. In addition, if you are unable to generate a stable cell line with a given receptor, it is still possible to run a transient HTS on the LumiLux. You can make a workable cell line in just days and work with aequorin technology, rather than spending 3 6 months generating a stable cell line. 384-well Aequorin Data Using Low Responding Transient Cell Line Receptor X in RH7777 Cells Receptor Y Stable CHO Cell Line FILE FROM DB NOT USABLE RH7777 cell responses in 384-well format transiently expressing aequorin and a GPCR. Stable CHO aequorin cell line responses in 384-well microplates are used as controls in high-throughput screening campaigns. Transient Transfection of Receptor X in RH7777 Cells Microplate Number EC 50 format of cells LumiLux 384-well 6, nm MicroLumat 96-well 50, nm Equivalent pharmacology generated for LumiLux (384-well) and a comparative PMT reader (96-well) with higher-fold induction and significant throughput improvements being generated with the LumiLux platform. (Data compliments of a large biotechnology company.) 7

8 Inside the LumiLux High-Impact Productivity Challenges Innovative cell stirrer enables screening with suspension cells The LumiLux integrates advanced fiber-optic imaging technology, state-of-the-art liquid handling and a unique cell stirrer that permits the use of suspension cells in uhts screens to provide an advanced standalone instrument with true walk-away capability. 8 Tipping plate and cell stirrer

9 Instrument Solves Cellular Screening 9

10 Comparison of the Different Methodologies for Running Aequorin Assays If you prefer to work with adherent or pre-dispensed cells, you can still work with the aequorin technology using the LumiLux platform. In the data shown below, a typical aequorin assay has been carried out using suspension cells, cells that have been plated for 24 hours prior to the assay, and cells that have been pre-dispensed into plates and left for 10 minutes prior to stimulation with compound. Comparison of Adherent and Suspension Cell Aequorin Assays Harvest cells Load with Coelenterazine Plate cells Aspirate Media Inject and read cells added to compound plates AEQUORIN SUSPENSION CELL ASSAY Inject cells into empty plate Inject compound and read AEQUORIN PRE-DISPENSED CELL ASSAY inject compound and read AEQUORIN ADHERENT CELL ASSAY Comparison of the Different Methodologies for Running Aequorin Assays Comparison of the Different Methodologies with Fluorescence Calcium Assays Robust Histamine responses are generated using luminescence and fluorescence methodology. The high S/B generated using the luminescence aequorin suspension technology enables increases in productivity, reduces screening and labor costs while maintaining assay quality. Agonist dose response curves using the different methodologies Antagonist dose response curves for the different assay methodologies Comparison of Histamine concentration response curves for CHO aequorin H1 cells in 384-well assays using different assay methodologies. Comparison of antagonist Triprolidine inhibition curves for CHO aequorin H1 cells in 384-well assay, for different assay methodologies. Assay Methodology Plate Z' S/B pec 50 Adherent (10,000 cells/well) Pre-dispensed (12,500 cell/well) Suspension (12,500 cell/well) Assay Methodology Plate Z' S/B pic 50 Adherent (10,000 cells/well) Pre-dispensed (12,500 cell/well) Suspension (12,500 cell/well)

11 Screening Results Summary Flexible acquisition modes improve throughput even further LumiLux has two acquisition modes: kinetic mode, which allows the time course of a flash luminescence to be measured; and integration mode, where a single image is taken for the whole of the luminescence peak. This functionality enables kinetic profiles to be viewed during assay development and the option to run high-throughput screens using a single integration, thus reducing data analysis. Integration mode is also suited for glow type luminescence. The bar chart to the right illustrates data that has been obtained in both modes. Comparison of Integration Times (20 sec v 400 ms) Comparison of Histamine positive control response (for S/B and Z') for CHO Aequorin H1 cells in a 384-well assay using kinetic and single read acquisition modes to measure light emission. Walk away and leave the LumiLux to screen, generating high performance data The LumiLux can be used to screen aequorin assays in both 384-well and 1536-well formats generating robust data for each day of a screening campaign. Data from the LumiLux running unattended, where the cell stirrer has been loaded with sufficient numbers of cells for a complete day s screening, is presented below. Day to day screening results, obtained across a campaign, are also presented. (Data compliments of a large biotechnology company.) Typical Screening Results Agonist and Antagonist Screen Campaign (384-Well) Data demonstrates cell and assay stability over a 6-hour period, supporting the robustness of the LumiLux Cellular Screening Platform as a fully automated uhts instrument and of aequorin technology. Data demonstrates assay stability over a 27-day period with Z' tracking around 0.7 throughput the combined agonist/antagonist screen. The number of plates increases during the initial setup to approximately 60 x 384-well plates being screened each day. (Data compliments of a large biotechnology company.) 11

12 Integrated Detection and Liquid Handling Robotics Provide precise, automated liquid handling functionality Walk away without worry from your sample preparation and screening workflow with LumiLux s automated liquid handling robotics. LumiLux s liquid handling functionality provides true walk-away automation, reducing hands-on time. The LumiLux s automated liquid handing system permits the workstation to perform multiple tasks simultaneously, offering a true parallel processing capability. The result is a breakthrough in sample processing speed over traditional serial operation systems. The workstation s liquid handling functions can operate in parallel with plate reading, performing operations such as compound and agonist/antagonist aspiration and dispensing, tip washing, etc. The system can aspirate more than one solution in the tip and inject them simultaneously. In addition, precision dispensing and diluting enable automation of mother-daughter replication from 96 up to 1536 wells, and serial dilutions in 96- and 384-well plates, adding key functionality for these labor intensive tasks. LumiLux is equipped with a 384-channel dispense module with a P30 disposable tip head. Using P30 tips, it has a pipetting volume range of µl, and using P10 tips, a pipetting range of µl. It achieves precision better than 5% CV across 384-well microplates when dispensing The LumiLux MultiPosition Dispense (MPD) module holds reagents for liquid handling, a 384-channel dispense head with integrated Gripper, and a positioner for accurate plate placement on the fiber-optic imager. 10, 20 or 30 µl and better than 10% CV across 1536 wells when dispensing 1, 2, 3 or 4 µl into 1536-well microplates. You can select and control variable liquid handling functions such as aspirate and dispense speeds, heights and volumes, tip wash, stackers, and de-lidding (to prevent condensation on plates and evaporation) through the system s WinPREP software. LumiLux enables complete automation of all assay steps via multiple deck positions and parallel processing. cell stirrer 384-channel dispense head unique optics Deck accessories for running all assay steps LumiLux Deck Design. The advanced and efficient deck design enables handling of up to 200 plates with no add-on robotics. It s bidirectional conveyor links the MPD, stacker and shuttle modules with specific application modules such as dispense, tipwash, plate de-lidding, barcode scanning, plate to waste, and reagent cartridges. All liquid handling components are 100% DMSO-compatible providing unrestricted use in pharmaceutical applications with DMSO-solubilized compounds. The combination of the conveyor with the Gripper tool increases productivity further through multi-tasking on the LumiLux. walkaway capability tipwash 12

13 Sophisticated Optics Deliver superior sensitivity for cellular assays LumiLux uses optics based on patented fiber-optic contact imaging technology with an ultra-cooled CCD camera, resulting in highly efficient light gathering and low crosstalk, to achieve a high level of sensitivity for a range of luminescence-based assays. LumiLux employs an ultra-sensitive, -80 C cooled, fiberoptic coupled CCD camera using a back-thinned CCD. The instrument can measure flash and glow luminescence in a variety of plate formats including 384-well and 1536-well plates. Contact imaging with fiber-optics produces highly efficient light transfer further increasing sensitivity. Fiberoptics eliminate the need for complicated lenses, and avoid lens effects such as vignetting, parallax, reflections and cross-talk (< 1% for 1536-well format). It even removes any requirement for focusing. The LumiLux optical system views sample plates from below, and is built into PerkinElmer s unique MultiPosition Dispense (MPD) module design. This allows inject-and-read capability for both kinetic and end-point assay applications. Fiber-optic taper images all wells simultaneously in glow assays and 384 wells simultaneously in flash luminescence assays. 13

14 Built-In Automation and Data Analysis Software Provide powerful tools for flash and glow luminescence assays, sample preparation and data analysis WinPREP Automation Software for liquid handling Automating liquid handling for your assays is simple with WinPREP software. Ready-to-go protocols for flash and glow luminescence assays, serial dilutions (384) and mother-daughter replications (384 to 384 and 384 to 1536) are pre-installed on the LumiLux. A visual application tool, WinPREP software is the industry standard in providing all the flexibility and power required for defining today s fully automated applications. AssayPro Data Analysis Software Our exclusive, easy-to-use AssayPro data analysis presentation software and database package lets you view data as it is collected, identifying hits and performing assay QC in real time. AssayPro also collects and stores raw data in an integrated ultra HTS database server. This facilitates detailed off-line analysis of large volumes of data by single or multiple users, and can even performing multiple data analyses simultaneously. AssayPro s off-line data analysis and graphical presentation tools enable generation of EC 50s, IC 50s, Max, Min, Area under the curve and more. AssayPro Analysis Protocol Map Define baseline and response windows Create multiple data analysis transforms in the Analysis Protocol Map Create assay specific plate maps and templates View data by plate Export your data to other applications Create doseresponse curves and calculate EC 50 14

15 With fully automated LumiLux, you just load and go! Run your screen in 3 simple steps Load cell stirrer with cells. Load stacker(s) with compound plates. Press go. Leave the LumiLux to screen! With LumiLux, you can run two screens a day, 6 hours each, 5 days a week!

16 Realize the high impact of LumiLux on your screening productivity! You can achieve unprecedented levels of productivity in cellular screening, virtually overnight. Invest in the LumiLux Cellular Screening Platform. You ll begin to impact your bottom line the first time you use it. To reach your PerkinElmer Account Representative, contact us at or locate your local sales office at Or visit our website at Worldwide Support Network People and applications supporting performance excellence PerkinElmer is recognized worldwide for advanced instrumentation systems and outstanding customer support. We are your partner in drug discovery and life science research providing instruments for scintillation, gamma, and microplate counting; quantitative imaging; and liquid handling. We are committed to providing the best instrument application support, field service and telephone support. We are proud of our tradition of working with laboratories to accelerate the pace of drug discovery and life science research. PerkinElmer Life and Analytical Sciences 710 Bridgeport Avenue Shelton, CT USA Phone: (800) or (+1) For a complete listing of our global offices, visit PerkinElmer, Inc. All rights reserved. The PerkinElmer logo and design are registered trademarks of PerkinElmer, Inc. AssayPro, Gripper and LumiLux are trademarks and WinPREP is a registered trademark of PerkinElmer, Inc. or its subsidiaries, in the United States and other countries. Windows is a registered trademark of Microsoft Corporation. All other trademarks not owned by PerkinElmer, Inc. or its subsidiaries that are depicted herein are the property of their respective owners. This device and operation thereof may be protected by one or more of the following patents: EP , US , US , GB Worldwide Patents Pending include EP PerkinElmer reserves the right to change this document at any time without notice and disclaims liability for editorial, pictorial or typographical errors _04 Printed in USA