Lum/nex ® Luminex 200™ User Manual for LDS Version 1.
© LUMINEX CORPORATION, 1998 - 2005. All rights reserved. No part of this publication may be reproduced, transmitted, transcribed, or translated into any language or computer language, in any form or by any means without prior express, written consent of: LUMINEX CORPORATION 12212 Technology Boulevard Austin, Texas 78727-6115 U.S.A. Voice: (512) 219-8020 Fax: (512) 219-5195 Technical Support U.S.
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strict compliance with the United States Food and Drug Administration and all applicable domestic and international laws and regulations, now and hereinafter enacted.
End-User License Agreement (EULA) for Luminex® Software This Luminex End-User License Agreement (“EULA”) is a legal agreement between you (either an individual or a single entity, also referred herein as “you”) the end-user and Luminex Corporation (“Luminex”) regarding the use of the Luminex software product identified above, which includes computer software and online or electronic documentation and may include associated media and printed materials (if any) (“SOFTWARE PRODUCT” or “SOFTWARE”).
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Contents Introduction 1-1 Principle of Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-1 Basic Concepts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-2 Fluidics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-2 Excitation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-3 xMAP Microspheres . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Luminex 200 User Manual for LDS Version 1.7 xMAP Technology Luminex XYP Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-4 Luminex SD System General . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-5 PC Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-5 Recommended Additional Equipment . . . . . . . . . . . . . . . . . . . . . . 3-5 Printer. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
xMAP Technology Contents Delete a Template . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-10 Multiplex Acquisition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-10 Menu Bar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-10 Toolbar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6-12 Main Control and Session Detail . . . . . . . . . . . . . . . . . . . . . . 6-12 Right-Click Menu . . . . .
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 Bead Detail Irregularities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Error Messages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Printing Errors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Verification. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Protocols 8-11 8-13 8-14 8-14 9-1 xMAP Microsphere Handling . . . . . . .
xMAP Technology Contents Luminex Sheath Delivery System C-1 Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . General . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Power . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Intended Use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Luminex 200 User Manual for LDS Version 1.7 vi xMAP Technology PN 89-00002-00-150 Rev.
1 Introduction The Luminex® 200™ system is a compact analysis unit consisting of a Luminex analyzer, the Luminex XY Platform (Luminex® XYP™), a PC, Luminex software, and reagents. The Luminex analyzer integrates optics, fluidics, and advanced signal processing in a 17 inch by 20 inch footprint. The Luminex XYP provides efficient handling of 96-well microtiter plates. The PC uses Luminex software to control the analyzer.
Luminex 200 User Manual for LDS Version 1.7 xMAP Technology its own subset, based on its fluorescent signature. In addition, the Luminex analyzer scans each microsphere for the presence of a reporter fluorescence that quantifies the assay at the microsphere’s surface. Microtiter plates with 96 wells must be compatible with the Luminex XYP plate holder.
xMAP Technology Introduction container and effectively removes residual sample within the tubing, valves, and probe. The second fluidics path is driven under positive air pressure and supplies sheath fluid to the cuvette. Excitation xMAP Microspheres Reporter Fluorochromes The excitation system in the Luminex analyzer uses two solid-state lasers.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 Quantum yield Fluorescence Compensation Repetitive Microsphere Measurements About this Manual 0.82 0.8 Fluorescent spillover occurs when an emission spectrum overlaps other emission criteria; therefore, selective filtering cannot occur. In most analyzers, emission spillover is corrected using a technique called compensation. Compensation involves subtraction of emission percentage from another emission signal.
xMAP Technology Introduction from the main menu bar, from the toolbar, and from menus that appear when you right-click an area of the screen. Technical Assistance You can find answers to frequently asked questions (FAQs) on our website: http://www.luminexcorp.com. From the Support menu at the top of the page, select FAQs. For assistance, contact Luminex Technical Support via email. Users may reach us at support@luminexcorp.com or supporteurope@luminexcorp.com. Users in the U.S.
Luminex 200 User Manual for LDS Version 1.7 1-6 xMAP Technology PN 89-00002-00-150 Rev.
2 Safety Your safety is important. Please do not perform procedures on your Luminex 200 system that are not specifically contained in this manual, unless you are directed to do so by Luminex Technical Support.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 Safety and Regulatory Symbols Symbol Description These symbols describe warnings, cautions, and general information used in the operation of this analyzer.
xMAP Technology Safety Danger: This message is used in cases where significant risk of serious injury or death is present. Safety Precautions Electrical Please read the following safety information before setting up or using the Luminex 200 system. A user should always be present during system operation. This system contains electrical, mechanical, and laser components which, if handled improperly, are potentially harmful.
Luminex 200 User Manual for LDS Version 1.7 xMAP Technology Figure 2-2. European Safety Requirements Label The Luminex analyzer complies with European Union (EU) safety requirements and, therefore, may be marketed in the Europe Single Market. The following voltage labels displays on the back of the Luminex analyzer: Figure 2-3. Luminex 200 Voltage Label (Coherent) This voltage label displays on the back of the Luminex XYP: Figure 2-4. Luminex XYP Voltage Label 2-4 PN 89-00002-00-150 Rev.
xMAP Technology Fluidics Warning: If the system is used to test biological samples, use your standard laboratory safety practices when handling system waste. Safety The Luminex analyzer contains fluidics. In the event of a fluid leak, turn off all power to the system and disconnect all power cords. Remember that the on/off switch is not a disconnect means - the power cord must be removed from the outlet. Contact Luminex Corporation for further information. You must manually monitor waste levels.
Luminex 200 User Manual for LDS Version 1.7 xMAP Technology Figure 2-6. Laser Caution Label Do not remove the analyzer cover. When performing routine maintenance, ensure that power to the analyzer is OFF and the power cord is disconnected. All laser apertures in the analyzer are contained within a protective housing. This label appears on the optics cover within the Luminex analyzer. Figure 2-7.
xMAP Technology Safety Figure 2-8. Avoid Exposure Label Caution: Use of controls or adjustments or performance of procedures other than those specified herein may result in hazardous radiation exposure. Caution: To avoid exposure to hazardous radiation, only perform procedures and adjustments as specified in this manual. Mechanical Warning: During operation, this system contains exposed, moving parts. Risk of personal injury is present. Observe all warnings and cautions.
Luminex 200 User Manual for LDS Version 1.7 xMAP Technology Heat Warning: The heater plate of the Luminex XYP may be hot and could cause personal injury if touched. Do not touch the heater plate. Blue Indicator Light Equipment Disposal The blue light above the sample arm simply indicates the on/off status of the instrument, and is harmless. The blue light emitting diode (LED) does not emit light in the UV spectrum.
3 System Overview Hardware The Luminex 200 for LDS Version 1.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 Required Laboratory Reagents Software The following items are necessary to run the Luminex 200 system. They are not provided by Luminex. • • • • Household bleach 70% isopropanol or 70% ethanol Mild detergent Distilled water Luminex Data Collector software provides complete control of the Luminex analyzer and performs real-time digital analysis of assays. The Luminex 200 system is preloaded with the Luminex software.
xMAP Technology Sensitivity System Overview • • Capacity The specifications below reflect minimum capacity values: • • • • • • • • • Luminex 200 Analyzer Specifications • • • • • • • • • • PN 89-00002-00-150 Rev. A Detect 1000 fluorochromes phycoerythrin (PE) per xMAP microsphere Reporter channel dynamic range: 3.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 Optics • AC inlet fuse: 3 Amp, 250 V~, fast acting • Reporter laser: 532 nm, nominal output 10-15 mW, maximum 500 mW, frequency-doubled diode; mode of operation, continuous wave (CW) Classification laser: 635 nm, 9.
xMAP Technology Luminex SD System General System Overview • • • • • • • • • PC Specifications Ambient temperature: 15°C to 30°C (59° to 86°F) Humidity: 20% to 80%, noncondensing Altitude: designed to operate at up to 2400m (7874 feet) above mean sea level Physical dimensions: 20 cm (8 inches) W x 30 cm (11.75 inches) D x 24.75 cm (9.
Luminex 200 User Manual for LDS Version 1.7 xMAP Technology and be UL listed, CSA certified, CE marked for nondomestic use when used internationally. Vortex Use VWR product number 58816-121: Speed range 0-3200 rpm or equivalent. Bath Sonicator Luminex System Overview Use Cole-Parmer® product number 08849-00: Operating frequency 55 kHz or equivalent. The Luminex analyzer consists of three subsystems: electronic, fluidic, and optical. In addition, several Luminex XYP system features are present.
xMAP Technology System Overview Luminex XYP Instrument Ventilation Filter The XYP instrument ventilation filter cleans the air that cools the internal parts of the Luminex XYP instrument. See Figure 3-1. Air Intake Access Door Power Switch Power Input Module XYP Instrument Ventilation Filter Communications Ports (DB9) XYP to PC Serial cable Analyzer Ventilation Filter Figure 3-1 Rear View of Luminex Analyzer PN 89-00002-00-150 Rev.
Luminex 200 User Manual for LDS Version 1.7 Fluidics Warning: During operation, this system contains exposed, moving parts which can cause puncture wounds. Risk of personal injury is present. Keep hands and fingers away from the sample probe. xMAP Technology Luminex XYP Sample Probe: A stainless steel sample probe that acquires the sample.. Cheminert® fitting: The fitting attaches the sample probe to the sample tubing. Disconnect this fitting when you remove the sample probe. See Figure 3-2.
xMAP Technology System Overview Left Door, Access to Service Panel Center Door, Access to Syringe Figure 3-3 Front View of Luminex analyzer Showing Access Doors Air filter and access door: A replaceable air intake filter cleans the air used to pressurize the sheath fluid. This filter is enclosed behind an access door located on the back of the Luminex analyzer. See Figure 3-4. Figure 3-4 Air Filter on Rear of Instrument PN 89-00002-00-150 Rev.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 Syringe: The syringe delivers a sample from the 96 well microtiter plate to the cuvette. It is located in the compartment behind the center door. 1 2 1. Syringe Seal 2. Syringe Figure 3-5 Syringe and Syringe Seal Sheath filter: The sheath filter removes particles greater than 10 microns in diameter from the sheath fluid. It is located behind the left access door.
xMAP Technology System Overview Sheath Fluid Connector (Blue) Air Connector (Green) Waste Connector (Orange) Figure 3-7 Left Side of Analyzer - Air and Fluid Connectors Sheath fluid container (not shown): Holds sheath fluid. For proper operation, place the Luminex SD system at the same level as the base of the Luminex XYP instrument. Do not put it on top of the Luminex 200 analyzer. If you are not using the SD system, sheath fluid levels must be monitored manually.
Luminex 200 User Manual for LDS Version 1.7 Optical Luminex XYP Reservoir Luminex XYP Heater Block 3 - 12 xMAP Technology The optical system contains the optics assembly and the excitation lasers. The optical assemblies do not require user adjustment. You can use 70% isopropanol or 70% ethanol, deionized water, and 10%-20% bleach solution in the reservoir for Luminex analyzer cleaning and maintenance procedures. Use the heater block as required by your assay procedures.
4 Installation Luminex 200 System Installation Note: The Luminex 200 System is too heavy for one person to lift alone. Two people should unpack and lift the system to the workbench. Ensure that the facility complies with all system and safety requirements. If you are using the optional Luminex SD, install it after completely installing the Luminex analyzer and the Luminex XYP. See the Luminex Sheath Delivery System appendix for instructions. To set up and start up the Luminex 200 system: 1.
Luminex 200 User Manual for LDS Version 1.7 xMAP Technology 5. Place the Luminex analyzer on top of the Luminex XYP. Do not plug it into the power outlet. 6. Without removing the Luminex analyzer alignment plate from the black knob, shift the analyzer so it is centered on the Luminex XYP. 7. Replace the short sample probe with the long sample probe as follows: Unsnap the light housing above the probe. Loosen the Cheminert fitting that holds the sample probe in place. Gently grasp the probe and push up.
xMAP Technology Luminex 200 Software Setup Installation After performing the Luminex 200 system assembly, ensure that the proper interface between the analyzer and PC is in place. To set up the software: 1. Turn on the Luminex analyzer, then turn on the PC. The Luminex software launches automatically. 2. Click Yes to accept the End-User License Agreement. If the system does not prompt you to accept this agreement, read the agreement shown after the title page of this manual.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 Adjusting the Sample Probe Height You must adjust the sample probe vertical height each time you install a sample probe, change the type or style of microtiter plate, move the instrument, or remove and replace the sample probe for cleaning. 1 2 3 4 5 1. Cheminert Fitting 4. Adjustment Slide 2. Sample Probe Holder 5. Sample Probe 3. Probe Adjustment Screw Figure 4-3.
xMAP Technology Installation For a filter bottom plate—stack three of the larger (5.08 mm diameter) alignment discs together and place them into well A1. For a half-volume plate with flat-bottom wells—stack two of the smaller (3.35 mm diameter) alignment discs together and place them into well A1. For a round-bottom (U-bottom) plate—stack two of the smaller (3.35 mm diameter) alignment discs together and place them into well A1. For a plate with conical wells—place one alignment sphere into well A1. 4.
Luminex 200 User Manual for LDS Version 1.7 Installing Software and Firmware xMAP Technology The Luminex 200 system arrives preloaded with the Data Collector software. It is possible that you may need to reload the software at some time. To reinstall the software: 1. Backup log files. Copy your template directory and the file named calibrationlog.mdb to a backup location. These files may be found in the Luminex setup directory. 2.
xMAP Technology Installation Time, Monday through Friday. Users outside of the U.S. and Canada can call us at +1 512-381-4397 between the hours of 7:00 a.m. and 7:00 p.m. Central Time, Monday through Friday. Users in Europe by can call us at +31-162408333 between the hours of 8:30 and 5:30, Central European Time, Monday through Friday. PN 89-00002-00-150 Rev.
Luminex 200 User Manual for LDS Version 1.7 4-8 xMAP Technology PN 89-00002-00-150 Rev.
5 System Startup This chapter describes how to start up your system. The topics included are: • • • Luminex Main Window Luminex Main Window Calibration Analyzer menu commands Use the Luminex software to start the system. To start the software and open the main window, double-click the Luminex Data Collector icon on the Windows desktop. Title Bar Menu Bar Tool Bar System Monitor Startup Display Status Bar Figure 5-1. Luminex Main Window PN 89-00002-00-150 Rev.
Luminex 200 User Manual for LDS Version 1.7 xMAP Technology The Luminex Main window has six components: Title bar, Menu bar, Tool bar, Startup display, Status bar, and System monitor. The Title bar shows the name of the software, and changes depending on what section of the software you are using. The Menu bar contains commands to set up the Luminex analyzer and perform data acquisition. See Chapter 6 for more detail.
xMAP Technology System Startup To add a new lot of xMAP classification calibration microspheres: 1. Click Calibrate. The Start Calibration dialog box opens. Figure 5-2. Start Calibration Dialog Box 2. Click New in the Classification Channel box. The Add New Classification Calibrator dialog box opens. Figure 5-3. Add New Classification Calibrator Dialog Box 3. Enter the information into the dialog box, using the values on the Certificate of Quality that came with your xMAP calibration microspheres.
Luminex 200 User Manual for LDS Version 1.7 Add New Reporter Channel Calibration Lots xMAP Technology Before you perform initial calibration for the analyzer, and whenever you receive a new supply of xMAP Calibration microspheres, enter the product and lot numbers and the calibration target values. To add a new lot of xMAP reporter calibration microspheres: 1. Click Calibrate. The Start Calibration dialog box opens. Figure 5-4. Start Calibration Dialog Box 2. Click New in the reporter channel area.
xMAP Technology Calibrate the Luminex Analyzer System Startup In addition to calibrating the analyzer after installation, you can enhance optimal performance by calibrating the instrument daily as part of your startup routine and whenever the d Cal Temp temperature shown on the system monitor panel indicates a change of more than 3 degrees Celsius (°C). To calibrate the Luminex analyzer: 1. Turn on the Luminex analyzer. Click Warmup on the main screen, then click OK on the dialog box that appears.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 7. Fill the reservoir with a solution of 70% isopropanol or 70% ethanol. Click Retract on the toolbar to retract the plate holder. 8. Click Alcohol Flush, then click OK. The alcohol flush removes air from the fluidics. This takes about 5 minutes. 9. Click Eject on the toolbar and replace the alcohol in the reservoir with distilled water. Click Retract on the toolbar to retract the plate holder. 10. Click Calibrate on the main screen.
xMAP Technology System Startup The progress bar shows the calibration progress, which should take less than two minutes. If the status changes to “Sample Empty” before calibration completes, a “Calibration Failed” message appears. Click OK and restart the process. When calibration finishes for the classification microspheres, a “Calibration Succeeded” message appears. The settings and any comment you enter are logged into the calibration report. 15. Click OK in the confirmation dialog box.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 Figure 5-9. Print Dialog Box 3. Check the box in front of the report type you want. You can check both boxes to see both kinds of calibration report. 4. Click OK to print the calibration report, or Preview to see the report on your screen. Figure 5-10 shows an example of a calibration log. Figure 5-11 shows an example of calibration detail report. Figure 5-10.
xMAP Technology System Startup more) in the gain value for a channel from one calibration to the next, the difference may indicate a problem with the instrument. It is normal to see a value of -1 for RP1 gain for classification calibrations. You also see gain values of -1 listed for DD, CL1, and CL2 for reporter calibrations. Figure 5-11. Luminex Calibration Detail Report The calibration detail report is often used by Luminex field service personnel when performing system maintenance.
Luminex 200 User Manual for LDS Version 1.7 xMAP Technology if a Luminex XYP instrument is added to the system. See page 4-3 for more information. Sanitize: Decontaminates the sample lines and cuvette after biohazard contact. Use a 10%-20% bleach solution to sanitize the system after running biohazardous samples. See page 7-3 for the sanitizing procedure. Prime: Removes air pockets from the fluidic pathways. Use this operation after system installation and following replacement of the sheath fluid.
xMAP Technology System Startup recommend that you do not cancel warmup. Your test results will be accurate only if the system is completely warmed up before you begin acquisition. A complete warmup cycle is critical for accurate calibration. XYP Retract: Retracts the plate into the Luminex XYP slot. Enable Heater: Warms the Luminex XYP heater block. PN 89-00002-00-150 Rev.
Luminex 200 User Manual for LDS Version 1.7 5 - 12 xMAP Technology PN 89-00002-00-150 Rev.
6 Data Acquisition The Luminex analyzer uses Luminex Data Collector software to acquire data. This chapter describes the following topics: • • • • • • Options User tab Options Sessions Templates Multiplex Acquisition Data Collection Reports Before you can acquire data, you must set up some of the different options available. Click Options on the toolbar to open the Options dialog box. The Options dialog box has these tabs: User, Acquisition, Advanced, Setup XY, and Info.
Luminex 200 User Manual for LDS Version 1.7 xMAP Technology Sound effects: Enables sounds during system operation. Clear recent file list: Removes the list of recent files from the File menu. Do this each time you want to clear the list of recent files. Show project settings dialog on new session: Opens the project settings dialog box when you start a new session. You can also click Settings on the toolbar to open the settings dialog box.
xMAP Technology Acquisition tab Data Acquisition On this tab you can select the default folder for your session files. If you want a folder other than the one shown, click Browse and select a different folder. Figure 6-3. Options Dialog Box - Acquisition Tab Advanced tab Use the Advanced Tab to set properties in the Luminex Data Collector software. You can select multiple commands in this tab. Figure 6-4. Options Dialog Box - Advanced Tab Show Dot Plot.
Luminex 200 User Manual for LDS Version 1.7 xMAP Technology Show APD Voltages. This is a Field Service option. Do not select this option unless directed by Technical Support Show Project Properties. This is a Field Service option. Do not select this option unless directed by Technical Support. View Calibration Detail. Shows the detail screen while calibrating the instrument. Allow FileMode. Enables use to replay previously run sessions. Enable Liquid Handler. This is a Field Service option.
xMAP Technology Warning: The heater plate is hot when in use and may cause personal injury. Do not touch the heater plate. Allow the heater block to cool after use before touching it. Data Acquisition To use the heater, check Enabled in the Heater area. Use the Set Point control to select a temperature between 35°C and 60°C (95°F to 140°F). To maintain heater temperature between sessions, check Hold Temperature between sessions checkbox.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 Create a Session Using Default Settings To create a session using default settings: 1. On the Luminex main window, click New Session. The Settings dialog box opens. The values shown in the General tab are recommended settings. Figure 6-7. Settings Dialog Box - General Tab 2. Enter a name for the session in the Description field. Enter your name in the Operator field. Modify other fields as necessary.
xMAP Technology Data Acquisition 2. Select the template that you want to use for this session and click OK. If the template already contains a beadmap (or MAP), the template defaults to that MAP. If you have not yet selected a beadmap, this Choose Bead Map dialog box opens: Figure 6-9. Choose Bead Map Dialog Box Select the MAP you want to use and click OK. The 100 Region, 50 Region, and 25 Region MAPs are the standard MAPs for multiple analyte data collection.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 Open and Add to an Existing Session To open an existing session and add to it: 1. Click Open on the toolbar. The Open an Existing Session dialog box opens. Figure 6-11. Open an Existing Session Dialog Box 2. Double-click the session folder that contains the session that you want to use. The session information displays in the session group box. Click OK. The Multiplex Acquisition window opens. Figure 6-12.
xMAP Technology Data Acquisition 3. Use the settings shown or modify them, then highlight the first vacant sample. 4. Click Start to collect the new data; it is added to the established session. Open an Existing Session and Save it as a New Session To open an existing session and save it as a new session: 1. Click Open on the toolbar. 2. Choose the session folder and click OK. The established session appears. 3. Click File on the main menu bar and select Save As, then rename your session.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 2. Make desired setting changes. See “Session Settings” on page 614. 3. Click Edit on the main menu bar and select Create Template. Supply the same name as the loaded template. 4. When prompted to overwrite an existing template, select Yes. 5. Close the session and select No when asked to save changes to the current session. Delete a Template To delete a template: 1. Click Tools on the main menu bar and select Manage Templates. 2.
xMAP Technology Data Acquisition Open. Opens an existing session Close Session. Closes the current session. Prompts you to save the session before closing. Save. Saves the current session. Save As. Saves the current session under a new name. Print. Opens the print dialog box. You have the option of printing a results table, calibration log, bar graph, histogram, and dot plot. Print Setup. Opens the Print Setup dialog box. Use the settings in this dialog box to choose printer options. Exit.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 Help Contents. Opens the online help. Search for Help On. Opens a searchable index for the online help. Luminex on the Web. Opens a web browser window and navigates to the Luminex website. About Luminex Data Collector. Provides information about the Data Collector software. Toolbar The toolbar gives easy access to important commands of the main menu. Each of the commands are listed in the previous section. Figure 6-14.
xMAP Technology Data Acquisition Clear All: Clears the screen of all data and graphs. This function deletes all associated files. A confirmation dialog displays when you click this button to make sure that you intend to delete the table, graphs, and the data files for the session. Autosize: Adjusts column widths to fit the data and header sizes. Statistic: This dropdown list offers a choice of statistic types displayed in the table. Changing the statistic type does not affect collected or recorded data.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 Right-Click Menu Other commands appear on the right-click menu. To open this menu, right-click any sample cell. Figure 6-16. Right-Click Menu Load Data: Displays the data, including histogram and events information, of a selected (highlighted) sample. Start/Cancel: Begins or ends data acquisition. Delete Data: Deletes the data from a selected sample row. Copy: Copies selected information from the results table to the Windows clipboard.
xMAP Technology Data Acquisition We recommend using at least 100 events per microsphere set. This guarantees that each bead set selected acquires 100 events before completing sample acquisition. If each selected bead set does not acquire this number of events, you see a warning that not enough bead events were acquired. You can choose to keep the data or disregard it. Because of this option, when selecting bead sets on the Bead Set tab, you should choose only the bead sets present in your sample.
Luminex 200 User Manual for LDS Version 1.7 Bead Set tab xMAP Technology On the Bead Set tab you can select and name the appropriate microsphere sets. Select only microsphere sets that are present in your samples. Figure 6-18. Settings Dialog Box - Bead Set Tab Available: Lists available bead sets for the default beadmap (or MAP) selected in Options. See page 6-2 for more information. If the bead set you need is not on the Available list, click Options on the toolbar and select a different default MAP.
xMAP Technology Histogram Data Acquisition The histogram is located in the lower-left corner of the screen. The histogram provides a graphical display of the real-time data collection. An example of a histogram follows: Figure 6-19. Histogram Example The Y axis of the graph displays collected events. To autoscale the Y axis: Click the up-and-down arrow button shown just above the graph. To manually set the Y axis scale: 1. Right-click anywhere in the histogram to open the right-click menu.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 2. Click X axis, then click on the desired parameter name. If you select reporter 1 for the X axis parameter, the histogram displays reporter-generated intensity data. The central collected wavelength is 575 nm. With classification 1 or classification 2, peaks are based on the emission wavelengths of the classification dyes inside the microspheres; the central collected wavelengths are 658 nm and 712 nm, respectively.
xMAP Technology Data Acquisition Use the Show Bead options to set the histogram to show events for only one bead set, all gated events, or all events. There are five buttons to the upper right of the histogram: Auto scale: Automatically adjusts the maximum number of events shown on the Y axis. Click during acquisition to readjust the Y axis scale. Zoom: Enlarges a specific area on the histogram. Use the cursor and left mouse button to adjust the graph’s range.
Luminex 200 User Manual for LDS Version 1.7 xMAP Technology for later assays or sessions. By increasing this voltage setting, you may gain sensitivity; however, you may also compromise range. Decreasing this value has the opposite effect. To return to the calibrated RP1 setting, click Edit on the main menu bar and select Reset to Calibrated Settings. We recommend that you use the calibrated RP1 value. Bar Graph/Dot Plot Display You can toggle between the bar graph and dot plot displays.
xMAP Technology Data Acquisition density is indicated by contrasting colors. A decaying dot plot shows the 100 most recent events acquired by the Luminex analyzer. Figure 6-26. Dot Plot Display In the upper right corner of the frame, four buttons let you control the display: Count/Result: Switches the display between count and result. This is either a count of the number of events collected for the bead ID, or an RP1 intensity result. The result is the median value for the events.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 the beads are hitting with respect to their region. The beads should hit within the white region shown. The colored dots show where the beads actually hit. Figure 6-27. Bead Details Box This display can be useful in troubleshooting problems. See the “Troubleshooting” chapter on page 8-1 for examples. If the beads do not register properly during sample analysis, recalibrate the Luminex analyzer.
xMAP Technology Data Acquisition The message at the top reflects the current status of the Luminex analyzer. Ready: Indicates that the Luminex analyzer is ready to collect data. Pressure: Pressure normally displays during a routine pressure check at the beginning of each run. However, if the pressure message remains for more than 1.5 minutes, a lack of pressure exists in the system. Check for the following problems: • • • • Low sheath fluid Loose hose connections Loose cap on sheath fluid bottle.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 The system monitor section also shows information about temperature, pressure, voltage, and events. Air pressure generally runs a little higher than sheath pressure. It is normal to see a drop in pressure if several minutes elapse between sample runs. Information in the calibration box of the systems monitor, which appears when no session is open, is shown for the last successful calibration.
xMAP Technology Data Acquisition 5. In the results table of the new session, highlight the sample where you want to begin collecting data. 6. Click Start. You can see the progress of data collection on the histogram and the bar graph. Make sure that the Single box remains unchecked for continued sample acquisition. 7. After the analyzer finishes collecting data, select Save on the File menu. The analyzer saves your data in the session folder you selected when you created the session.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 9. Remove the microtiter plate from the plate holder. 10. Click Retract from the toolbar to retract the plate holder. Run a Range of Samples You can run a specific range of samples, as demonstrated in the highlighted samples shown below. 1 2 3 4 5 6 7 8 9 10 11 12 A B C D E F G H Figure 6-31. Sample Plate with Row of Samples To run a range of samples on a plate, follow these steps: 1. Create the session for this run. 2.
xMAP Technology Data Acquisition 9. Remove the microtiter plate from the plate holder. 10. Click Retract from the toolbar to retract the plate holder. Run a Single Sample To run a single sample: 1. Click Eject from the toolbar to eject the plate holder. 2. Place a 96-well microtiter plate on the plate holder. 3. Click Retract from the toolbar to retract the plate holder. 4. On the results table part of the acquisition screen, select the sample that you want to read. 5.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 To interrupt sample acquisition between sample runs: 1. Click Cancel. Sampling stops after you click Cancel. 2. Click OK to continue acquiring the next sample. Reports You can print the results table or histogram from any session. To print reports: Note: The analyzer does not support printing with the Windows NT operating system. 1. Load the desired data. 2. Click File on the main menu bar and select Print. The Print dialog box opens.
xMAP Technology Data Acquisition 6. In the Manufacturers List, click Generic. In Printers list, click Generic/Text Only. 7. Click OK throughout the rest of the windows. 8. You can now open report pages in Notepad, then print them to this printer. PN 89-00002-00-150 Rev.
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7 Maintenance and Cleaning Warning: When analyzing potentially infectious biological samples on the Luminex analyzer, follow proper biosafety procedures. These safety precautions should also be taken when cleaning or maintaining the Luminex analyzer. Proper maintenance and cleaning of the Luminex analyzer should be performed in order to preserve the analyzer’s longevity and reliability.The maintenance log at the end of this chapter will help you to keep track of when maintenance procedures are performed.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 Startup Note: There is NO sensor to warn you of low sheath fluid levels. Fill the sheath fluid container before the level reaches the sheath outlet line. There is NO indicator light to warn you of high waste volume. Empty the waste container each time you fill the sheath container. Perform the entire startup procedure on a daily basis. 1. Turn on the analyzer and PC. 2. Click Warmup. 3.
xMAP Technology Check Fluid Levels Maintenance and Cleaning Check the sheath and waste fluids daily, more often if you run several samples. Replace the sheath fluid and empty the waste container as needed. You must manually monitor the sheath fluid and waste container levels. Release the Luminex analyzer’s pressure by removing the lid from the sheath fluid container. Add sheath fluid, filling the sheath fluid container to just below the air intake. Discard waste by appropriate means.
Luminex 200 User Manual for LDS Version 1.7 Clean Accessible Surfaces xMAP Technology Clean the exterior surfaces of the Luminex 200 system on a monthly basis, or more often if you are running biological samples. 1. Turn off the power switch on the rear of the Luminex analyzer. 2. Unplug the analyzer power cord from the wall source. 3. Wipe all exterior surfaces with mild germicidal detergent, followed by a 10%-20% bleach solution. 4.
xMAP Technology Replace Air Intake Filters Note: Hold on to the tubing! Do not allow the tubing to fall inside the analyzer. Maintenance and Cleaning You should replace the filters on the Luminex analyzer and Luminex XYP every six months. To replace the air intake filter on the Luminex analyzer: 1. Turn off the power switch on the rear of the Luminex analyzer, then unplug the power cord from the wall source. 2.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 Figure 7-2. Back of Luminex XYP, Removing Air Filter 3. Replace the filter and replace the screen. 4. Plug in and turn on the Luminex XYP. Replace Syringe Seal Replace the syringe plunger seal every 6 months. 1. Turn off and unplug the Luminex analyzer. 2. Open the center door on the front of the Luminex analyzer. Warning: The syringe arm does not deactivate when changing the plunger; injury could result if the system is not unplugged. 3.
xMAP Technology Maintenance and Cleaning 1 2 1. Syringe Seal 2. Syringe Figure 7-3. Front of Luminex 200 Analyzer, Access Door Opened Clean Luminex Analyzer Ventilation Filter Check the Luminex analyzer ventilation filter every 6 months. Clean the filter only when soiled. 1. Turn off the Luminex analyzer at the power switch. 2. Unplug the power cord from the wall source. 3. On the bottom of the unit on the left side, push the filter out toward the right side. 4.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 3. Connect the new sheath filter, matching up the color-coded fittings. The arrow on the sheath filter should point up. 4. Reconnect the sheath fluid bottle. 5. Prime the instrument 5 times to remove any air introduced into the system. Figure 7-4. Sheath Fluid Filter Change Sheath Quick-disconnect O-rings Change the sheath quick-disconnect o-rings as necessary. 1. Turn off the Luminex analyzer at the power switch. 2.
xMAP Technology Change Fuses Maintenance and Cleaning To replace fuses on the Luminex analyzer or the Luminex XYP instrument. 1. Turn off the power switch on both the Luminex analyzer and the Luminex XYP. DANGER! To avoid serious injury or death by electric shock, you must turn off the system and unplug it from the wall outlet. 2. Unplug the power cord from the analyzer or instrument, depending on which fuses you are changing. 3. Use a small, flathead screwdriver to open the power module door.
Luminex 200 User Manual for LDS Version 1.7 Storing the Analyzer xMAP Technology To prepare the system for storage: 1. Run a Sanitize command with 10% to 20% household bleach solution. 2. Run a Sanitize command with distilled water. 3. Run four Wash commands with distilled water. 4. Remove the sample probe from the instrument, flush with distilled water from the narrow end out through the larger end, replace it in the sample arm and wrap the end with Parafilm.
xMAP Technology Decontaminating the Luminex Analyzer for Return Shipment Maintenance and Cleaning Luminex Technical Support will give you a Return Material Authorization (RMA) number if they direct you to return the system. They will explain how to return the system according to Luminex procedures. The accessible surfaces and the internal fluidics system must be sanitized and decontaminated before returning the analyzer. This is particularly important when biohazardous samples have been run.
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Luminex 200 Maintenance Log Month(s): ________________________________________Year:_______________ Procedure: To use this log, fill in the month(s) and year above. This form is designed to record information for four weeks. Make photocopies as necessary. Fill in the dates for which this log is used in line one of the table below. For each item listed at the left, add your initials under each date you perform the item.
8 Troubleshooting Troubleshooting procedures help users isolate, identify, and remedy problems with the Luminex analyzer and Luminex XYP instrument. This chapter does not troubleshoot problems with the PC. For help with PC problems, please call Dell Technical Support at 800-6249896. To troubleshoot a problem, start by selecting a general symptom Next, identify the possible problem and remedy it with one of the solutions listed. This troubleshooting chapter supplies information for the following topics.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 For assistance, contact Luminex Technical Support via email. Users may reach us at support@luminexcorp.com or supporteurope@luminexcorp.com. Users in the U.S. and Canada can call us at 1-877-785-BEAD (1877-785-2323) between the hours of 7:00 a.m. and 7:00 p.m. Central Time, Monday through Friday. Users outside of the U.S. and Canada can call us at +1 512-381-4397 between the hours of 7:00 a.m. and 7:00 p.m. Central Time, Monday through Friday.
xMAP Technology Troubleshooting • Symptom Instrument control, sample acquisition, session uploading, and start, stop and pause features. Possible Problem Solution PC cannot initialize communication with analyzer. The communications cable is unplugged. Check the communications cable connections. “Bad Link” error message There is a bad link between the PC, analyzer, and/or XYP. Ensure that the instruments have power.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 Pressure too High Fluid Leaks Symptom Pressure too low The compressor does not engage. Run a Prime command. If you do not hear the compressor turn on, call Technical Support. The Cheminert Fitting is loose. Ensure that the fitting connects tightly above the sample probe, below the blue light. Fluid leaks in the system. See Fluid Leak on page 8-4. The sheath bottle has an air leak.
xMAP Technology Fluid leaking from front of analyzer Sample Probe Symptom Troubleshooting The sample 3-way valve is faulty. Contact Technical Support. The syringe seal leaks. Replace the syringe seal. The syringe valve leaks. Hand-tighten the syringe connection (silver knob) to the white syringe valve. Run a Prime. If leaks continue, contact Technical Support. Problems with the sample probe can lead to fluid leaks and pressurization problems, as well as inhibit sample acquisition.
Luminex 200 User Manual for LDS Version 1.7 xMAP Technology Sample arm stuck in the down position The sample probe height is too low, or the path to the well is blocked. DO NOT turn off Luminex XYP instrument. 1. Remove the blue light housing on the Luminex analyzer. 2. Unscrew the tubing connector connecting the sample tube to the top of the sample arm. The system monitor changes from “Busy” to “Running” and sample acquisition continues.
xMAP Technology Troubleshooting The sample probe is bent. Remove the sample probe from the Luminex analyzer. Roll it on a flat surface. If it does not roll smoothly, replace it with a new sample probe. There is a bad link between the PC and the analyzer. Verify that all instruments turned on and all communications cables are in place on back of instruments. The calibration microspheres are not fully suspended. Vortex the calibration vials to resuspend the microspheres.
Luminex 200 User Manual for LDS Version 1.7 Classification channel calibrates, but Reporter channel fails. 8-8 xMAP Technology The calibration microspheres are not fully suspended. Vortex the calibration vials to resuspend the microspheres. The wrong calibration lot was chosen in the Calibration Setup window. In the Main Window of the Luminex Data Collector software, click Calibrate.
xMAP Technology Zero events collected during calibration Troubleshooting Beads were left in the system after Classification calibration. Perform an Alcohol Flush followed by a Wash command. There is a problem with fluid levels. Check the fluid levels in the sheath and waste containers. Verify that both bottles are tightly connected to the instrument. Check that the waste bottle cap is vented. Acquisition Problems Symptom Acquisition fails or slows PN 89-00002-00-150 Rev.
Luminex 200 User Manual for LDS Version 1.7 Slow or Unsuccessful Sample Acquisition 8 - 10 xMAP Technology The wrong wells are selected in the Setup XY tab. On the Options Menu of the Luminex Data Collector software, click Options. When the Options window opens, click on the Setup XY tab. In the Default Sample Locations group box, make sure that the correct corresponding wells are chosen from the drop-down list. The air pressure is out of range. See Pressurization on page 8-3.
xMAP Technology Bead Detail Irregularities Troubleshooting You can diagnose many problems by using the histogram described on page 6-17 or the Bead details box described on page 6-20. A normal bead detail display is shown below. It depicts a tight bead population within a white region. Figure 8-1. Normal Bead Detail The histogram for the bead detail above looks like this: Figure 8-2. Normal Histogram Symptom PN 89-00002-00-150 Rev.
Luminex 200 User Manual for LDS Version 1.7 xMAP Technology Calibration microspheres classify too high You may be using photobleached calibration microspheres. Replace the microspheres with a fresh batch. To avoid photobleaching, protect your microspheres from light. xMAP microspheres hit toward the lower right of the region You may be using photobleached xMAP microspheres. Replace microspheres with a fresh batch. To avoid photobleaching, protect your microspheres from light.
xMAP Technology Troubleshooting Error Messages Error Type Unable to calculate temperature compensation. Possible Cause The delta Cal temperature is out of range. Solution Recalibrate the system. If message continues after two more attempts at calibration, contact Technical Support. Syringe pump error “A CAN The syringe seal is loose. protocol (bad frame) error occurred during step 2 of the stop sample sequence.
Luminex 200 User Manual for LDS Version 1.7 Sheath Fluid Error The sheath fluid bottle is empty. xMAP Technology Refill the sheath bottle, then run several Prime commands until you see fluid flowing into the waste bottle. If the message does not appear after refilling sheath fluid, disconnect the bottom fitting of the sheath filter shown on page page 7-7. Plug the waste bottle into this fitting and leave the sheath bottle, sheath, and air lines connected normally. The analyzer is not pressurizing.
xMAP Technology Troubleshooting 1. Click Warmup on the toolbar. Wait 30 minutes for the warmup to complete. 2. On the Analyzer menu, click Prime. Prime the system twice. 3. On the Analyzer menu, click Backflush. Perform this command twice. 4. From the Analyzer menu, click Alcohol flush. Use 70% isopropanol or 70% ethanol to flush the system. 5. Calibrate the system. 6. Vortex the xMAP calibration (CAL1) and xMAP reporter (CAL2) calibration bead vials.
Luminex 200 User Manual for LDS Version 1.7 xMAP Technology from the Luminex 200 system, copy the files to a floppy disk and send them from another computer. 16. Technical Support uses these files to verify that your system runs as it should. 8 - 16 PN 89-00002-00-150 Rev.
9 Protocols Updates or additions to these protocols are posted on the Luminex website at http://www.luminexcorp.com.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 Note: Failure to perform this procedure can result in decreased amounts of xMAP microspheres recovered in future sample aliquots. 3. Sonicate the vial in a water bath sonicator to disperse the microsphere pellet. 4. Vortex the vial immediately prior to dispensing. Larger 4 mL and 16 ml vials should be placed on a rotator for a minimum of 15 minutes for complete resuspension.
xMAP Technology Enumeration of xMAP Microsphere Suspensions Protocols Luminex xMAP microspheres are provided at standard concentrations. The xMAP microsphere yield after a coupling process may be less than the starting concentration due to loss during wash steps. The xMAP microsphere loss can vary according to operator technique, coupled reactant properties, and scale of coupling. It is not advisable to construct an assay without defining the concentration of the coupled microsphere suspension.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 xMAP Microsphere Stability and Storage xMAP microspheres are light sensitive and should be protected from light during all stages of storage and usage. In addition, freezing conditions and organic solvents should be avoided. Two-Step Carbodiimide Coupling of Protein to xMAP Carboxylated Microspheres Introduction Equipment Materials Use the protocols included here as a general starting point for developing assays.
xMAP Technology Protocols • • • Preparation Sulfo-NHS: N-Hydroxysulfosuccinimide sodium salt, Pierce Chemicals EDC: 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride, Pierce Chemicals Protein for coupling (See Technical note 2) 1. Allow all reagents to warm to room temperature. 2. Dilute the protein stock with COUPLING BUFFER to a concentration of 25 - 250 µg/ml and a volume of at least 500 µl (See Technical note 2). 3.
Luminex 200 User Manual for LDS Version 1.7 xMAP Technology 11. Incubate the suspension for 20 ± 2 minutes in the dark at room temperature. 12. Centrifuge the activated xMAP microspheres for 1 minute at 8,000 × g or higher. Aspirate the supernatant (See Technical note 3). 13. Add 500 µL of COUPLING BUFFER to the activated xMAP microspheres and vortex. Centrifuge the activated xMAP microspheres for 1 minute at 8,000 × g or higher. Aspirate the supernatant. 14.
xMAP Technology Protocols amines. If any of these agents exist in the protein preparation, remove them through dialysis or chromatography. 3. During activation, centrifuged xMAP microspheres may form a sheet-like layer on the side of the microfuge tube. This is normal. 4. For best results, we recommend using microcentrifuge tubes from USA Scientific, Inc., catalog number 1415-2500. To order, call 1-800-LAB-TIPS.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 • Preparation SDS: Sodium Dodecyl Sulfate (0.1% w/v) 1. Allow all reagents to warm to room temperature. 2. Resuspend the amino-substituted oligonucleotide to 1.0 mM in sterile, deionized water. Procedure 1. Centrifuge the xMAP microsphere stock for 1 minute at a minimum of 8,000 x g. 2. Disperse the pellet with sonication, and vortex the container for 20 seconds. 3. Dispense 5.0 × 106 of the xMAP microspheres from the stock tube into a 1.
xMAP Technology Protocols 16. Centrifuge the xMAP microspheres at 8,000 × g or higher for 1 minute. 17. Remove the supernatant, being careful not to disturb the pellet. 18. Resuspend the xMAP microspheres in 100 µl of TE (10mM Tris, 1mM EDTA, pH 8.0). 19. Enumerate the xMAP microsphere preparation. 20. Store the preparation at 2oC - 8°C. Protect it from light. Technical notes 1. Oligonucleotides must be synthesized with a 5’ or 3’ amino group.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 Materials • • • • Micropipetters (1 µl - 1000 µl) Microcentrifuge Timer Rotator • xMAP LumAvidin-modified microspheres—LIMIT EXPOSURE TO LIGHT! Biotin-conjugated molecule Microcentrifuge tubes: 1.5 ml, polypropylene REACTION BUFFER: Phosphate Buffered Saline (pH 7.3 ± 0.1), Bovine Serum Albumin (10 mg/ml) BLOCKING/STORAGE BUFFER: Phosphate Buffered Saline (pH 7.3 ± 0.1), Tween 20 (0.02% v/v), Bovine Serum Albumin (1 mg/ml), Sodium Azide (0.
xMAP Technology Protocols 9. Add 500 µl of BLOCKING/STORAGE BUFFER to the microspheres and vortex. Centrifuge the tube for 60 seconds. Aspirate the supernatant. 10. Repeat step 9. 11. Resuspend the microspheres in the desired volume of BLOCKING/STORAGE BUFFER. 12. Enumerate the microsphere preparation. 13. Store the preparation at 2oC - 8°C. Protect it from light. Technical notes 1. The concentration of Biotin-conjugated molecules is provided as a guide for assay development.
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A Glossary aggregated microspheres Two or more xMAP microspheres that have adhered together. Alexa 532 Fluorochrome used for reporter channel with Ex/Em=531/554. (product of Molecular Probes, Inc.) ambient temperature The temperature of the surrounding environment. analyte A substance that is detected through assay analysis. Each test or unique xMAP microsphere type will test for a specific analyte. assay A quantitative measurement of a particular compound.
Luminex 200 User Manual for LDS Version 1.7 xMAP Technology calibrators xMAP microspheres used to normalize the settings for the reporter channel, both classification channels, and the doublet discriminator channel for the Luminex analyzer. carboxylated microspheres An xMAP microsphere set that has been chemically modified so that its surfaces are covered with carboxyl functional groups (COOH) that allow covalent biomolecule attachment. CL1 Refers to dyes embedded in the xMAP microsphere.
xMAP Technology emission spectrum emission spectrum Wavelength range that an excited fluorochrome emits when its electrons fall from a higher to a lower energy state. Expressed in nanometers (nm). event Occurs when the signal processor determines that a particle is being observed. Ex/Em A common way of reporting a molecule’s excitation and emission wavelengths (for example, Ex/Em = 488/520). excitation spectrum Wavelength range that excites a molecule’s electrons to a higher energy state.
Luminex 200 User Manual for LDS Version 1.7 xMAP Technology xMAP microsphere set Luminex multi-analyte microspheres containing a unique mixture of two distinctly-colored fluorochromes to distinguish them from other multi-analyte microspheres. laser Light Amplification by Stimulated Emission of Radiation (laser). This highly purified source of light is an efficient way to excite fluorochrome electrons.
xMAP Technology reporter reporter A molecule (or combination of molecules) with a specific range of excitation and emission wavelengths that are used to identify or quantify an analyte. Examples of acceptable reporters for the Luminex analyzer are Phycoerythrin, Alexa 532, and Cyanine 3. reporter channel (RP or RP1) A specific range of wavelengths that includes the emission wavelength of a designated reporter molecule.
Luminex 200 User Manual for LDS Version 1.7 A-6 xMAP Technology PN 89-00002-00-150 Rev.
B Product Numbers Note: These part numbers are subject to change without notification. Hardware PN 89-00002-00-150 Rev. A Air filter (accessed through back of analyzer) CN-0001-01 Air intake filter (on bottom of analyzer) CN-0002-01 Serial cable CN-0005-01 Computer (Dell® PC) CN-PC01-01 Fuses, 2 Amp CN-0019-01 Heater block for XYP CN-0017-01 Luminex 200 (Analyzer) with XYP (Optional Luminex SD and PC or laptop) LX200-LDS1.7 Luminex 200 User Manual for LDS Version 1.
xMAP Technology Luminex 200 User Manual for LDS Version 1.
xMAP Technology xMAP development microspheres (carboxylated) 1, 4, and 16 mL L100-CDEV1-[size code of 01, 04, or 16] xMAP development microspheres (LumAvidin), 1 mL L100-LDEV1 xMAP carboxylated microspheres 1, 4, and 16 mL L100-C1[region number]-[size code of 01, 04, or 16] for example, to order 1 mL of region 34 microspheres, use product number L100-C134-01 xMAP (LumAvidin) microspheres, 1 mL L100-L1[region number]-[size code] for example, to order 1 mL of region 22 microspheres, use product number
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C Luminex Sheath Delivery System Overview Use this appendix to install, use, and maintain the Luminex Sheath Delivery System (Luminex SD™). Specifications General • • • • • • • Power Intended Use • • Ambient temperature: 15°C to 30°C (59° to 86°F) Humidity: 20% to 80%, noncondensing Altitude: designed to operate at up to 2400m (7874 feet) above mean sea level Physical dimensions: 20 cm (8 inches) W x 30 cm (11.75 inches) D x 24.75 cm (9.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 WARRANTY RIGHTS PROVIDED BY LUMINEX AND/OR ITS AUTHORIZED PARTNER. Hardware The Luminex SD system includes: • • • • • • • Luminex Sheath Delivery device Waste line tubing (orange fitting) Sheath fluid line (blue fitting) Air line (green fitting) Sheath fluid intake line (white fitting) Power cord Screwdriver 1. 2. 3. 4. 5. 6. 7. 1. Power and Fault Indicators 2.Prime Button 3. Regulator Adjust 4.Air In Connector (green) 5.
xMAP Technology Sheath Delivery System Setup Installation Luminex Sheath Delivery System When adjusting the Luminex 200 regulator, turn it clockwise as far as it will go to the fully closed position, then turn it back 1/2 of a turn. Be careful not to force the regulator past the fully closed position. To install the Luminex SD system: 1. Locate the sheath fluid and waste bottles that are connected to the Luminex analyzer. Check that the sheath fluid bottle is filled to just below the air intake valve.
xMAP Technology Luminex 200 User Manual for LDS Version 1.7 Note: Ensure that the sheath fluid box is at least three feet below the system, otherwise the Luminex SD system will overfill. • Connect the other end of the air line to the air connector on the left side of the Luminex analyzer (green fitting). • If you are using the Luminex SD system waste line, connect the waste line tubing to the waste connector on the left side of the Luminex analyzer (orange fitting).
xMAP Technology Luminex Sheath Delivery System 10. In the Luminex Data Collector Main window, click Prime. During this prime cycle, use a screwdriver to adjust the regulator on the front of the Luminex SD system. See Figure C-2. Adjust it until the sheath pressure displayed in the System Monitor reads the same as the sheath pressure you recorded in step 4. The system should stabilize at this sheath pressure. The air pressure should be the same as you recorded in step 4, within 0.1 psi.
Luminex 200 User Manual for LDS Version 1.7 Maintenance and Cleaning xMAP Technology Perform proper maintenance and cleaning of the Luminex SD system to preserve the instrument’s longevity and reliability. Regularly, as required. Replace the sheath fluid as required. Weekly. Visually inspect the Luminex SD system and its connections for leaks. If you see a leak, turn off the power to the Luminex SD system and contact Luminex Corporation. Monthly. Turn off the power to the Luminex SD system.
xMAP Technology Luminex Sheath Delivery System Filter. If the filter attached to the sheath intake line becomes clogged from extended use, an alarm will sound even though the bulk sheath container is not empty. If this happens, contact Luminex Technical Support for a replacement filter. Malfunction. If the alarm sounds even though the bulk sheath container has fluid and the sheath filter is in good condition, the system is reporting a malfunction. If this happens, please call Technical Support.
Luminex 200 User Manual for LDS Version 1.7 Labels C-8 xMAP Technology The following labels are attached to the back of the Luminex SD. PN 89-00002-00-150 Rev.
Index Numerics 100 region bead map 6-2 25 region bead map 6-2 50 region bead map 6-2 A access doors 3-8 accuracy/precision specifications 3-2 Acquisition tab 6-3 additional equipment 3-5 bath sonicator 3-6 uninterruptible power supply (UPS) 3-5 vortex 3-6 adjust sampling probe, vertical height 4-4 adjusting sampling probe 4-4 adjusting vertical height 7-9 Advanced tab 6-3 air connector 3-10 air filter 3-9 maintenance 7-5 product number B-1 air intake filter product number B-1 replacing 7-5 XYP 3-7 air pres
xMAP Technology Luminex 200 User Manual for LDS Version 1.
xMAP Technology heater temperature 6-22 height adjustment locking screw 4-4 high voltage 6-22, 6-24 histogram 6-17 buttons 6-19 changing X-axis 6-17 display 6-10 gating 6-18 printing 6-28 scaling Y-axis 6-17 I Info tab 6-4 installation 4-1 instrument operation 2-2 intended use 2-1 intended use, Luminex SD C-1 L label 2-4 laser 1-3 laser apertures 2-6 laser radiation 2-6 laser warnings 2-5 leaks 2-5, 7-3 load data 6-14 location 2-6 log report 5-8 lot number for calibrators 5-4 Lots, add new classification
xMAP Technology Luminex 200 User Manual for LDS Version 1.
xMAP Technology accuracy/precision 3-2 capacity 3-3 classification laser 3-4 electronics 3-4 fluidics 3-4 Luminex 100 IS 3-4, 3-6 PC 3-5 reporter laser 3-4 sensitivity 3-3 speed 3-2 vortex 3-6 specifications, Luminex SD C-1 specifications, Luminex XYP 3-4 speed specifications 3-2 spillover, fluorescent 1-4 standby 6-23 start 6-13, 6-14 startup procedure 7-2 status 6-22 status bar 6-10, 6-24 support 1-5, 8-1 symbols 2-2 syringe 3-10 syringe cylinder product number B-2 syringe seal maintenance 7-6 product nu
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