® xPONENT for ® FLEXMAP 3D Software Version 4.
© Luminex Corporation, 2012. 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. LUMINEX CORPORATION 12212 Technology Boulevard Austin, Texas 78727-6115 U.S.A. Voice: (512) 219-8020 Fax: (512) 219-5195 xPONENT® for FLEXMAP 3D® Software Verison 4.2 User Manual 89-00002-00-399 Rev.
Standard Terms and Conditions for Use of Instrument Product By opening the packaging containing this instrument product ("Product") or by using such Product in any manner, you are consenting and agreeing to be bound by the following terms and conditions. You are also agreeing that the following terms and conditions constitute a legally valid and binding contract that is enforceable against you.
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End-User License Agreement (EULA) for Luminex® xPONENT® 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 xPONENT software product provided to you above, which includes computer SOFTWARE and online or electronic documentation and may include associated media and printed materials (if any) (“SOFTWARE”).
SOFTWARE on additional computers, as agreed to in writing with Luminex or its authorized reseller, for purposes of performing ancillary tasks (i.e. preparing templates/ protocols, performing further analysis or re-running previous data), provided such computers are at a single location and are NOT connected with a Luminex instrument.
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Table of Contents Chapter 1 Introduction .......................................................................................................1 Safety Precautions .......................................................................................................................................1 Elements of the Software .............................................................................................................................1 Home Page ...............................................
Importing a Batch ................................................................................................................................49 Exporting a Batch ................................................................................................................................49 Editing a Batch ....................................................................................................................................50 Deleting a Batch ..............................................
Commands and Routines ..........................................................................................................................97 Creating a New Routine ......................................................................................................................99 Editing a Routine ...............................................................................................................................100 Deleting a Routine .....................................................
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Chapter 1: Introduction Safety Precautions DANGER: Samples and waste fluid can contain biohazardous infectious agents. Handle them at Biosafety level 2, as recommended for any potentially infectious human serum or blood specimen in the DCE/NIH manual, Biosafety in Microbiological and Biomedical Laboratories, 1984. CAUTION: Although beads do not contain hazardous or carcinogenic components at toxic levels, they can be toxic if swallowed. In addition, contact with acids liberates toxic gases.
The Home page displays a Welcome message, batch creation buttons, Daily Activities shortcuts, and the Installed Protocols list. Return to the Home page at any time by clicking Home in the Navigation toolbar. This page contains the following: Click to Create a new Batch using the highlighted Protocol below - Creates a new batch from a new protocol. This allows you to create a new protocol while you are creating the batch.
• System Initialization - Opens the routine specified in the Maintenance Options on the Admin page. • Shutdown - Opens the System Shutdown command in the Auto Maint tab on the Maintenance page. • Probe and Heater - Opens the Probe and Heater tab on the Maintenance page. Sys Info - Opens the System Info tab of the Maintenance page. Reports - Opens the Reports tab of the Results page. Screen Elements This section shows screen elements and the terms used in this book to describe them.
Right-Click Menu Certain sections of the software such as tables, lists, and text boxes have right-click option menus. Menus are different depending upon the item you right-clicked. • Apply Dilution Down - sets the dilution levels for the sample. • Print All - Prints all sections or cells of the item. • Print Selection - Prints only the selected section or cell. • Import - Imports a file. • Export Grid Contents - Opens a File Dialog dialog box.
Command Display - Displays the following: • The command currently running. • The system state (i.e. running, idle, etc.). • Date and time. Progress - Displays a bar graph showing the progress of the current command or routine; if the command or routine is finished, it displays a full progress bar and the command status as Complete. Pause - Pauses the system after the current command completes. Pause does not stop the system in the middle of running a command.
To Go to Expanded Help Adjust the sample probe height Home > Probe and Heater Adjusting the Sample Probe Initialize the system Home > System Initialization Running the System Initialization Routine Run an assay Home > Create a new Batch Create a New Batch from a New from a new protocol, or Home Protocol > Create a new Batch using the highlighted protocol below Create a New Batch from an Existing Protocol Analyze Results > Saved Batches Performing Analysis Print reports Results > Reports Repo
• Serial Number • Firmware Version • XYP Heater Temperature • Sheath Pressure • Calibration/Verification Status • Last F3DCAL1 Calibration • Last F3DeCAL1 Calibration • Last F3DCAL2 Calibration • Last F3DCAL3 Calibration • Last F3DVER1 Verification • Last F3DeVER1 Verification • Last F3DVER2 Verification • Last Fluidics Test • Delta Calibration Temp • System Temperature • DD Temperature • CL1 Temperature • CL2 Temperature • CL3 Temperature • CL1 Voltage • CL2 Voltage • CL3 Voltage • DD Voltage • RP1 Voltage
Basic Procedures StartingxPONENT® • On the PC desktop, click the Luminex® xPONENT® icon, or click Start > All Programs > Luminex > xPONENT > Luminex xPONENT. • If you have a trial license, contact Luminex® Technical Support to obtain a full license, or click OK in the dialog box to continue. • If this is the first time you have started the software, the User License Agreement may display. Read the license agreement. Select I accept the terms of this license agreement, then click OK.
CAUTION: Use of this software by untrained personnel can result in inaccurate data and test results. Users of xPONENT® must read the documentation thoroughly before operating the software. 1. On the System Login tab, type your user ID. 2. If you are using a secure version of the software, type your password. The Home page opens. 3. From the Home page, open the Admin page. Open System Setup tab. NOTE: This allows you to define the System Initialization routine.
4. From the Maintenance options drop-down menu you can define the daily startup routine. Select the check box if you want to Allow running the calibration or verification if the instrument is not warmed up. 5. You can set how often the system will run the calibration routine. Initial Startup When you turn on the system for the first time, perform the following procedures: 1. Adjusting the Sample Probe Height 2.
NOTE: If using a strip-well plate, ensure that a strip is present in the selected well location, prior to calibration. 5. Place the plate on the plate holder with A1 in the top left corner. Place the off-plate reagent block on the plate carrier. Place a well-strip (provided with the Calibration and the Performance Verification kits) in column S1 of the block. NOTE: Make sure there is no liquid in the plate or the off plate reagent block. 6. Click Retract to retract the plate carrier. 7.
WARNING: Correct sample probe height is critical to successful sample acquisition and calibration. Problems with the sample probe can lead to fluid leaks and inhibit sample acquisition. CAUTION: Ensure that the probe height is set correctly before calibrating the system. Revive After Storage Routine NOTE: The Revive After Storage routine is necessary when the system has been idle for more than a week. After you have adjusted the sample probe height, run the Revive After Storage (Luminex) routine. 1.
NOTE: See the Defining the System Initialization section for more information. Calibrator xMAP beads are used to normalize the settings for the reporter channel, all classification channels, and the doublet discriminator channel. Verification xMAP beads are used to verify calibration and optical integrity of the system. If the system is not fully calibrated, a warning message opens. Once calibrated, the values remain until you recalibrate.
3. Click Exit to exit the application. Using Online Help English-language help is available at all times while you are using xPONENT®. To display online help for the page or tab in which you are currently working, click the blue “i” icon at the upper-right of the xPONENT® window. This opens a help window with information specific to that page or tab. To display system-level help, click the blue question mark at the top of the xPONENT® window, then click Contents and Index.
Luminex® Support Viewing the Luminex® Website Additional information is available on the Luminex® website. FAQs are available at http:// www.luminexcorp.com/Support/index.htm. You can access the Technical Support Website using a user name and password at https:// esupport.luminexcorp.com/OA_HTML/jtflogin.jsp. Contacting Technical Support Luminex® Technical Support representatives are ready to help you. If the question or problem relates to materials from the assay kit, contact the kit provider directly.
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Chapter 2: Running Assays with FLEXMAP 3D® General Guidelines WARNING: Modifying or deleting xPONENT® system files can cause degradation of system performance. Repair modified or deleted xPONENT® system files by uninstalling and re-installing the xPONENT® software. Luminex ®recommends that you contact Luminex® Technical Support before uninstalling and reinstalling xPONENT®. WARNING: Using unauthorized third-party software with xPONENT® software can result in corruption or failure of the xPONENT® software.
Biological Samples CAUTION: Human and animal samples may contain biohazardous infectious agents. Where exposure to potentially biohazardous material—including aerosol—exists, follow appropriate biosafety procedures and use personal protective equipment such as gloves, gowns, laboratory coats, face shields, or mask and eye protection. Use ventilation devices. Observe all local, state, and federal biohazard handling regulations when disposing of biohazardous waste material.
• Maximum well volume plate = The maximum volume capacity of the wells in a selected 96 and 384-well microtiter plate. NOTE: This sample volume information is for a 96 and 384-well plate. Plates Follow the IVD assay manufacturer's instructions for use on the selection of plates. If not specified, follow these guidelines when choosing plates: • When using uncovered plates, use black opaque plates to reduce photobleaching.
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Chapter 3: Samples Page Samples Page Overview Samples > Samples Use this tab for the following: Create New Samples - Click to create a new sample. Protocol - Displays the protocol selected in the Samples tab. If xPONENT® has a Laboratory Information System (LIS) license enabled, any sample details provided by the LIS also appear in the Sample list. Version - Displays the protocol version number.
Number of Samples - Displays the number of samples are in the protocol. NOTE: If a protocol is created using the same name and version as a previously deleted protocol, previous, or pending are relinked to the added protocol. Details - Displays the details of the selected protocol. Create Batch - Creates a new batch from the selected protocol. Creating New Samples Follow these steps to create a new sample list. 1. Open the Samples page.
2. In the Sample Lists section, select the protocol you are using for the sample list, then click Create New Samples. The Create Sample tab opens. 3. In the ID box, type the sample ID. WARNING: You will not warned if you create a sample ID that is the same as another. 4. Type a patient first name in the First name box (optional). 5. Type a patient last name in the Last name box (optional). 6. Type any comments (optional). 7. Click Save to add the sample to the Sample list. 8.
Editing a Sample List 1. Open the Samples page.
2. In the Samples list section, choose the protocol you want to edit, then click Details. The Edit Samples subtab opens. 3. Click a sample, then use the Move arrows to move it up or down in the sample list, changing the order in which they will be acquired. 4. To add a new sample to the list, click New, then perform the following steps: a. In the ID box, type the sample ID. b. Type a patient first name in the First name box if desired c. Type a patient last name in the Last name box if desired. d.
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Chapter 4: Batches Page Batches Page Functionality Batches > Batches Options on the Batches tab on the Batches page are: • Create a New Batch from an existing Protocol • Create a New Batch from a new Protocol • Create a New Multi-Batch 27
NOTE: These tabs (except New MultiBatch) are sequential. You must complete each screen in a specific order. The Pending Batches list displays the name of the protocol used with the batch, the protocol version, date, and status for each pending batch. The following buttons only appear if the pending batches have data: Reacquire - Reacquires the selected batch. WARNING: Performing this action may result in varying results due to decreased bead counts, potential carry over effects, and timesensitive reagents.
Using the Batches Page 1. Open the Batches page. 2. Click one of the following: • Create a New Batch from an Existing Protocol • Create a New Batch from a New Protocol - if you select this option, you can save the protocol and/or stds/ctrls information. • Create a New Multi-Batch 3. Type the batch name in the Batch Name box. 4. Type an optional description of the batch in the Enter Optional Description box. 5. If you are creating a batch from an existing protocol, select the protocol in the list.
NOTE: If the batch spans more than one plate, the tray ejects automatically when all defined wells have been acquired. A dialog box opens, prompting you to insert the next plate. Protocol Subtab Batches > Batches > Protocol Use this tab to name a batch, type a batch description, select a protocol, and view active reagents. This tab contains the following: Batch Name/Description - Used to name and describe a batch.
Use this tab to apply a kit or lot to the batch. This tab contains the following: Apply Std/Ctrl Kit - Opens the Select Std/Ctrl Kit dialog box. The dialog box displays the Std/Ctrl Kit Lot #, Std/Ctrl Kit Name, Expiration, and Manufacturer for the kit. Select a Std/Ctrl kit from the list and then click OK to close the dialog box. The kit information will display in the boxes to the right of the Apply Std/Ctrl Kit button. The selected kit must be associated with the same analyte names.
Apply Std Lot - Opens the Select Lot dialog box. Select a lot from the list and then click OK to apply the lot. Apply Values - Applies a value across or down the Reagent, Name, Lot #, Expiration, and Analyte fields. Type a value in these fields by double-clicking on them, and then using one of the two Apply Values arrows to apply that value down or across the list of analytes. Dilution - Contains the following dilution options: • 1:2 - Halves the standard from each previous iteration.
Cancel - Returns to the Batches tab. Back - Returns to the previous tab. Next - Continues to the Plate Layout tab. Plate Layout Subtab Batches > Batches > Plate Layout Use this tab to define commands that apply to one or more wells. You can also define offplate and maintenance commands. This tab contains the following: Plate Image - This is a representation of the plate. Each well is displayed as a circle on the grid.
Replicate Count - Defines a quantity of replicate sets from one to nine. NOTE: Replicate count selection must be made before adding a well command. Grouping - Selects the sequence in which the replicates are laid out in plate wells. NOTE: Grouping selection must be made before adding a well command. The options are: • 123123123. . . Lays out one of each replicate set at a time in numerical order. • 111222333. . . Lays out all the replicates in a set before moving on to the next set in numerical order.
an error when attempting to run a batch on a system where the routine does not exist. Clicking either Pre Batch Routine or Post Batch Routine opens the Commands and Routines dialog box, where you can select the command or routine you want before or after running the batch. Clicking Add after selecting a well opens the same box for you to select a command or routine for that well. Clicking Delete after selecting a well deletes any commands or routines associated with that well.
5. Define the settings in the Acquisition Settings section. These are Volume, XY Heater (enable/disable and set temperature), Plate Name, and enable/disable Sample Wash. NOTE: Final washes are required for proper analysis. If you are not performing a final wash step on your plate before acquisition on the FLEXMAP 3D® instrument, enable Sample Wash here. This automatically washes each sample.
16. Click Single Step to acquire the first well, then pause acquisition. 17. Click Run Batch to start acquisition, or Save to save the batch for a later time. You can also save the protocol and/or standard and control information by clicking Save Prtcl. NOTE: If the batch spans more than one plate, the tray ejects automatically when all defined wells have been acquired. A dialog box displays prompting you to insert the next plate.
CAUTION: Acquiring data before the heater has reached the proper temperature can compromise test results. • Timeout - Check the Enabled box to enable the Timeout. • Bead Type - Check the Bead Type box. From the drop down menu you can select MicroPlex or MagPlex. • Plate Name - Select the plate name from the drop down menu. Select Current 96-well plate, Current 384-well plate, or a plate that you have created. • DD Gating (Doublet Discriminator) - Use this to set the minimum and maximum DD Gating values.
• Number of Standards - Click to type the number of standards for the protocol. • Number of Controls - Click to type the number of controls for the protocol. • Fit of all Standards - The standard curve is determined by using each individual standard replicate when calculating the standard curve. For example, if you run duplicates of a 7-point standard curve, the software will calculate the standard curve by using 14-points.
Default Analysis - The default analysis changes based on the Analysis Type selected in the Settings tab. You can change the analysis settings for all analytes by clicking Change if this button is enabled in this tab. This displays the Analysis Settings dialog box. FIGURE 1. Analysis Settings Dialog Box If you selected Quantitative on the Settings tab, the default analysis formula is 5P Weighted.
Selected Analytes List - Selected analytes appear in a list on the right side of the analyte grid. This list includes the following information: • Name - The name of the analyte. Click and type to rename the analyte. • Analysis - To change the type of analysis for an analyte, click this field to open the Analysis Settings dialog box and select another analysis from the list. • Select a method from the Method list. • If necessary, select a weight type from the Weight Type list.
Use this tab to apply a kit or lot to the batch. This tab contains the following: • Apply Std/Ctrl Kit - Opens the Select Std/Ctrl Kit dialog box. The dialog box displays the Std/Ctrl Kit Lot #, Std/Ctrl Kit Name, Expiration, and Manufacturer for the kit. Select a Std/Ctrl kit from the list and then click OK to close the dialog box. The kit information will display in the boxes to the right of the Apply Std/Ctrl Kit button. The selected kit must be associated with the same analyte names.
• 1:2 - Halves the standard from each previous iteration. • 1:10 (Log) - Computes a value of one-tenth of the standard from each previous iteration. • 1/2 Log - Creates a 1:3.16 dilution, or half of each 1:10 (Log) from each previous iteration. • Apply Dilution - Applies the dilution selected in the Dilution list. NOTE: The Dilution list and Apply Dilution button are displayed only if a quantitative analysis has been selected. NOTE: You can also manually type a number to set your own dilution factor.
Use this tab to define commands that apply to one or more wells. You can also define offplate and maintenance commands. This tab contains the following: • Plate Image - This is a representation of the plate. Each well is displayed as a circle on the grid. Well commands appear in the appropriate circles as you assign them to wells on the plate. • Command Sequence - Contains the command sequence for the active plate.
• 123123123. . . Lays out one of each replicate set at a time in numerical order. • 111222333. . . Lays out all the replicates in a set before moving on to the next set in numerical order. You can assign the following well commands. Each command is associated with a color.
• Plate Navigation - Displays a smaller plate image for the current batch. • Single Step - Instructs the system to acquire one well and then stop. Use this to ensure the system is working correctly before you run the whole batch. • Off Plate Area - Designates an alternate location for maintenance commands in the Commands and Sequence list. • Save Prtcl - Opens the Save Protocol dialog box to save the protocol and/or kit. • Select Save Protocol and/or Save Std/Ctrl Kit to save the protocol and/or kit.
Multi-Batch - Lists pending batches selected for the multi-batch. The list includes name and “Start at” well. Plate Layout - Opens the Multi-batch Report dialog box that contains: • Page - Use these arrows to scroll through the report pages. • Zoom - Select from the list to change the report magnification. • Print - Prints report. • Save - Saves report. • Close - Closes report dialog box.
2. Once you've selected a batch click Ok. 3. On the next screen, select a well location for the next batch and click Add.
4. Once the batches are created, click Run. After creating a multi-batch, you can save it to the Select Pending Batch list. When saved to this list, the protocol appears as “Multibatch.” To save a Multi-Batch: 1. Create a new Multi-Batch. 2. Select a pending batch. 3. Type the name for the Multi-Batch Name field. 4. Click Save. You are returned to the Batches page, and the multi-batch is added to the pending batches list.
4. Navigate to the location to which you want to save the file, then click Save. 5. Click OK in the Export Batch dialog box. NOTE: When exporting a large batch and including the LXB files, such as a batch with hundreds of wells or hundreds of analytes, the export process may take ten minutes or more. Editing a Batch 1. Open the Batches page. 2. Click the batch you want to edit, then click Edit. The Protocol tab opens. 3.
Chapter 5: Results Page Results Page Once data is collected in a batch, observation and analysis take place in the Results page. This page contains the following tabs: Current Batch - View statistics for the current run and progress per well. Saved Batches - View information about already processed batches and if necessary, replay them or recalculate their data. • From the Saved Batches tab, select the Open button located at the bottom right of the screen.
• Batch Audit • Patient Report • Protocol Reports • Protocol Settings • Plate Layout • Protocol Audit • Calibration and Verification Reports NOTE: Calibration and Verification Reports gives you Dates fields to select which dates to view the report. • All • F3DCAL1 • F3DeCAL1 • F3DCAL2 • F3DCaL3 • F3DVER1 • F3DeVER1 • F3DVER2 • Fluidics • Performance Verification Reports NOTE: Performance Verification Reports gives you Dates fields to select which type of report you want to view between specified dates.
Current Batch Tab Results > Current Batch Use this tab to view results, statistics, and log information related to the current batch, and to perform statistical analysis on batch results. This tab offers real-time monitoring of batch sampling during acquisition through a display of sample bead statistics and analytes, and dot plot data. The statistics available on this tab are intrawell bead statistics. They do not describe replicate well assay results.
The Current Batch tab has the following features: • Statistic - To view a particular statistic for analytes in a batch, select one of the options on the drop-down list. The statistical options displayed change according to the type of analysis. • Median (MFI) - The point is a data set where there are as many values occurring above it as there are below it. • Net MFI (Median Background) - Net MFI median background. • Count - Gated events, if gates are set.
• Analyte - Contains a list of analytes run in the batch. Select an analyte to view all statistics for that analyte. • Well(s) to View • Current Well - Displays the statistics of the well currently being displayed. (This changes to Displayed Well if viewing a batch using the Open button of the Saved Batches tab.) • Single Step - Instrument analyzes one well at a time. Click to turn function on or off. This is useful to run prior to running an entire batch to confirm that the system is set up correctly.
changed to visually inspect the effect. Only the visual display of the dot plot and histogram is affected for saved batches, not the acquired data. NOTE: Gate positions are dependant upon buffer composition. Any changes made to you assay buffer composition may result in a different optimal gate location.
• Date • Message • Code Log entries indicating warnings are highlighted ion yellow. Errors are highlighted in red. Other log entries are not highlighted. Click the maximize button to maximize the log. The log expands to fill the whole window. Click minimize to return to the standard size. • Save Image - Opens a Save As dialog box to save a screen capture. • Details - Opens the Results tab, enabling more analysis and results. • Progress - Click to display real-time progress of the well acquisition.
• 3D BeadMap - The main display is the 3D BeadMap. The display shows the CL1, CL2, and CL3 axes. Left-click and drag your mouse to move the display for better viewing of a particular section. Right-click anywhere in the display to select from the following viewing options.
• Linear • Display Position • 3D View • CL1/CL3 Plane • CL2/CL3 Plane • CL1/CL2 Plane • View • All Events • Gated Events • Show Cursors • Show Region Labels • Show 3D Region View Box • 3D Region View - This view in the upper right of the window displays a small 3D representation of the regions. The three axes are shown in the bottom left corner of the display and can be manipulated by left-clicking and dragging the mouse. Move the slider on the right to zoom in or out.
Use this tab to open a batch that has been run and view its details, and to export, approve, or replay a batch. Select a Completed Batch and click Open to view the batch details. The Saved Batches tab has 4 subtabs: • Click the Results subtab to view statistical information about the batch. • Click the Settings subtab to view the batch settings report. • Click the Log subtab to view a log for the activity that occurred during acquisition of the selected batch.
This dialog box lets you choose the saved batches you want to display in the Completed Batches list, based on options you select or clear in these check boxes: • Batch Name • Protocol • Batch Status • Lot ID • Kit ID • Analyte • Sample ID • First Name • Last Name • User ID • Date • Reset - Clears all check boxes. • OK - Closes the dialog box and applies any changes you made. • Cancel - Closes the dialog box and cancels any changes that you made.
• Plate Layout - Opens the Report dialog box, which contains the Batch Plate Layout Report. • Approve - Opens the Batch Approval Confirmation dialog box to approve the selected batch. Only approved batches can be transmitted to the LIS. If your software is licensed for LIS use, you can transmit batches to the LIS from the Sample Results tab. After you have approved a batch, the status of the batch changes to Approved in the Complete Batches list.
• Recalculate data - Reanalyze the batch results using only the batch MFI values. The batch MFI values are recalculated using the analysis settings and plate layout selected in the new recalculated batch or protocol. Settings such as bead type, volume, timeout, XY heater, and report gain have no effect on the replayed results. Because only the MFI values are reanalyzed, no data is displayed in the dot plot. • OK - Saves your changes. • Cancel - Cancels your changes and closes the box.
This subtab displays the following features: There are three maximize buttons in this window, one for each major pane. Click the appropriate one to maximize the pane. After clicking, the clicked button becomes a minimize button. Click minimize to return the pane to its standard size. This tab has the following features: • Statistic - To view a particular statistic for analytes in a batch. NOTE: The statistics displayed change according to the analysis type selected.
• Trimmed%CV of microspheres • Trimmed Standard Deviation • Trimmed Peak • %CV of Replicates • % Recovery • Expected Result - The known or expected test result value for a standard or control. • Control Range - Low - The lowest value for an assay control used to determine pass/ fail criteria for an assay. • Control Range - High - The highest value for an assay control used to determine pass/ fail criteria for an assay. • Normalized New Median • Units • Analyte - Contains a list of analytes run in the batch.
• Close - Closes the batch and reopens the Saved Batches tab. • Save - Saves the batch. This button only appears if a change has been made to the batch. • Analyze - Analyzes data that has been invalidated. This button only appears if you have clicked Invalidate on a portion of the data, or if you have clicked Validate on an invalidated portion of the data. This will analyze the data again with the invalidated standard removal.
Viewing Batch Settings 1. Open the Results page, then open the Saved Batches tab. 2. Click Saved Batches, then click the batch for which you want to view details. 3. Click Open, then click the Settings tab. 4. Click the left and right Page arrows to view the pages of the batch settings report. 5. Click Save to open the Save As dialog box. Navigate to the location where you want to save the batch settings report, and click Save.
• Export - Opens the Save As dialog box to save the batch log file. Select a location and click Save. • Close - Reopens the Saved Batches tab. Viewing Batch Logs 1. Open the Results page, then open the Saved Batches tab. 2. Click Saved Batches, then click the batch for which you want to view details. 3. Click Open. The Results tab opens. 4. Click Log to open the Log tab.
Viewing Sample Details 1. Open the Results page, then open the Saved Batches tab. 2. Click Saved Batches, then click the batch for which you want to view details. 3. Click Open, then click Sample Details. The Sample Details tab opens. If you are using an LIS licensed package of the software, click Transmit to transmit sample details to the LIS database. You can transmit either a single analyte per sample or the entire sample.
Others • User ID • Date • Reset • OK / Cancel • Clear - Click to turn off the filter. • Completed Samples - Displays Name, Protocol, Sample Count, Date, Status, and User information for each batch displayed in this list. • Transmit - Transmits a batch to a LIS if xPONENT® is connected to one. • Details - Opens the Sample Details tab to view sample results. Reports Tab Results > Reports Use this tab to view, generate, and print reports. Report and Type lists - Report lists the categories of reports.
• Data Interpretation • Batch Settings • Plate Layout • Batch Audit • Patient Report If you choose Data Interpretation, a Select Analysis section appears on the right and displays a list of the analytes in the batch. Select the analytes you want to include in the report. Select All to select all the analytes in the list. Select Clear to clear all the analytes in the list. A series of new buttons appears when you click Generate Report. • Analyte arrows - This feature is directly below the Report list.
Perfomance Verification Reports - Displays two report types. Choose either of the following: • Condensed • Detailed Select a report type, a date range from the Date list, and then click Generate to generate the report. System Log Reports - Displays a list of system log reports. Choose from the following selections: • All • Maintenance • Security • Warnings and Errors Select a system log report, a date from the Date list, and then click Generate to generate the report.
Chapter 6: Protocols Page Protocols Page Functionality Protocols > Protocols The Protocols page enables you to create or import a new protocol, or select an existing protocol from the Installed Protocols list.
• Name • Version • Manufacturer • Date Click the Create New Protocol button or click Stds/Ctrls to move to those pages. There are action buttons at the bottom of the page. Most of these buttons are not displayed unless you are working with a saved protocol. While you are creating a protocol, only Cancel and Next buttons are displayed.
8. Click Next. The Analytes tab opens. 9. Click the desired analyte (bead ID) in the numbered grid. Information about the analytes is displayed in a list on the right side of the grid. For an allele call analysis, you must select a group of two, three, or four analytes. 10. Click Group to group analytes for the allele call. The grouped analytes display in a list to the right. Select more analytes and then click Group again if you want to add another group to the analysis. 11.
NOTE: The protocol for a quantitative assay must have at least two or more standards. The number of controls are optional. 8. Click Next. The Analytes tab opens. 9. Click the desired analytes (bead ID) in the numbered analyte grid. Information about the analyte displays to the right side of the grid. 10. Click and type an analyte name in the Name column to the right of the analyte grid. 11. Click and type the desired unit of measurement in the Units box to the left of the Apply All button. 12.
• Check Mark as Intra-Well Normalization Bead if you want to use the analyte as a normalization bead. NOTE: The normalization bead is a microsphere set that is included in the assay as an internal control. It controls for sample variation and can be used to normalize data between samples in a run. • Check Use Threshold Ranges if you want to use a range for the analysis. • Click Add Range to setup the threshold range. • Type a name for the range in the Range Name field.
6. Define settings in the Analysis Settings section, selecting Qualitative as the analysis type. NOTE: The protocol for a qualitative assay contains a single standard. The number of controls are optional. 7. Click Next. The Analytes tab opens. 8. Click the desired analytes (bead IDs) in the numbered grid. Information about the analyte displays in a list on the right side of the grid. 9. To change the Default Analysis, click Change. The Analysis Settings dialog box opens. 10.
Exporting a Protocol 1. Open the Protocols page, then open the Protocols tab. 2. Select a protocol. 3. Click Export. The Save as dialog box opens. 4. Select a location to export the file to, and click Save. Importing a Protocol 1. Open the Protocols page, then open the Protocols tab. Click Import. 2. In the Open dialog box, navigate to the protocol file you want to import, then click Open. 3. The imported protocol displays in the Installed Protocols list. Adding a New Lot for Protocol 1.
3. If the protocol uses standards, type the appropriate information for each standard in the Assay Standard Information section. In each analyte column, type the expected concentration for the analyte. 4. Alternatively, click Apply Std/Ctrl Kit and select a lot from the Select Lot dialog box. Click OK to apply the lot. 5. If your batch uses controls, select Expected, Low, or High from the Show Value options. Use the Apply Values arrows to apply values down or across the range of analytes. 6.
3. Type the name of the kit in the Name box, the lot number in the Std/Ctrl Kit Lot# box, the expiration date using MM/DD/YYYY format in the Expiration box, and the manufacturer in the Manufacturer box. 4. Click Apply Std Lot if you want to apply a standard lot. The Select Lot dialog box opens. Click a lot and select OK. 5. Click Apply Ctrl Lot to apply a control lot. The Select Lot dialog box opens. Select a lot and click OK. 6.
Installed Protocols - Displays installed protocols. The list includes Name, Version, Manufacturer, and Date information for each protocol. New Std/Ctrl - Opens the Std/Ctrl Details tab. Plate Layout - Open the Report dialog box, which includes the Plate Layout Report. Delete - Opens the Delete Protocol dialog box to delete a protocol selected in the Installed Protocol list. Click Yes to delete the selected protocol. Import - Opens the Open dialog box to select a protocol (.lxt file) to import.
add at least 25 µl to the sample well in addition to the sample size. The default value is 45 µl. • XY heater - Select Enabled to enable the XY heater. In the box type the desired value in degrees Celsius. The temperature range is 35° C to 60° C. CAUTION: Acquiring data before the heater has reached the proper temperature can compromise test results. • Timeout - Check the Enabled box to enable the Timeout. • Bead Type - Check the Bead Type box. From the drop down menu you can select MicroPlex or MagPlex.
NOTE: Luminex recommends Fit of All Standards as the most accurate calculation of the curve fit. Based on a range of quantitative, numerical results, a threshold range can be applied to a quantitative analysis, for example, high, low, saturated, and expected. • Allele Call - Sets analysis for an allele call. Analytes must be put in groups of 2, 3, or 4. • Number of Standards - Click to type the number of standards for the protocol. Applies only to qualitative and quantitative analysis.
7. If you selected Lum Qual in the Formulas list, edit the values for each range under Negative, Low Positive, or High Positive. If you selected Adv Qual in the Formulas list, edit Negative, Low Positive, Moderate Positive, or Strong Positive. Select the check box in the Inclusive column to include the value in the range. If you clear the check box, the value will be one unit higher than the low value, and one unit lower than the high value. 8. Click Add Range if you want to add a new range.
12. Highlight a range and click Delete Range to delete the range. 13. Click OK to apply the new settings to the analyte first clicked, or Apply to All Analytes to apply them to all the analytes in the protocol. Plate Layout Subtab Protocols > Protocols > Plate Layout • Plate Image - This is a representation of the plate. Each well appears as a circle on the grid. Well commands appear in the appropriate circles as you assign them to wells on the plate.
• Replicate Count - Defines a quantity of replicate sets from one to nine. • Grouping - Selects the sequence in which the replicates are laid out in plate wells. The options are: • 123123123 - This displays one of each replicate set at a time in numerical order. • 111222333 - This displays all the replicates in a set before moving on to the next set in numerical order. You can assign the following well commands. Each command is associated with a color.
• Pre Batch Routine - Opens the Commands and Routines dialog box so you can select a pre-batch command or routine. • Plate Name - Specifies the plate you want to display in the plate image in the list. Add Plate adds a new plate to the batch, and Delete Plate deletes the plate highlighted in the list. Plate Name - This list displays the available plates to use for the selected routine or procedure. WARNING: When using multiple plates, use them in the proper order.
Use this tab to select or edit analytes used in the batch or protocol. This tab contains the following: This tab contains the following: • Analytes grid - A grid representing each analyte from 1 to 500. Select All selects all analytes, and Deselect All deselects all analytes. Click a numbered analyte to select it; click the analyte again to deselect it. You can also click and drag to select groups of analytes. Selected analytes are red. Deselected analytes are gray.
sets that are not in the sample. Therefore, choose only the bead sets present in your sample. • Apply All - Applies the information in the Units and Counts boxes to all analytes. • Total Count - Select Stop after bead count reaches: to stop acquisition when the bead count reaches a certain number determined by the user. Type the desired value in the box. The default value is 50. You can also specify the minimum allowable bead count per well that the xPONENT® software analyzes.
• Units - The unit of measurement you specified in the Unit box. Click this box to type a value for the analyte. • Count - Type the desired bead count for the analytes by clicking in the Count box. If each selected bead set does not acquire this number of events, a warning is added to the log that not enough bead events were acquired. • Region - Refers to the specific analyte selected. This is a number between 12 and 78.
After you select from this box, the Std/Ctrl Details tab opens so you can create a new lot or kit. • Installed Kits And Lots - Displays information about the kits and lots currently installed on the system. • Show - Click either Std/Ctrl Kits Only or All Lots to choose what to display. • Group lots from the same Std/Ctrl Kit together - Select to group lots from the same kit together.
Chapter 7: Maintenance Page Use the Maintenance page to maintain and calibrate your system.
This tab contains routines to initialize, verify, and maintain the analyzer. Automated Maintenance Options - Provides buttons for commonly used maintenance routines for the machine. Select from the following: • Calibration/Verification - Select this routine to use with the Calibration/Verification kit. • Performance Verification - Select this routine to use with the Performance Verification kit. • Fluidics Prep - Select this routine to use with Fluidics Prep.
Running Calibration and Verification Before running Calibration/Verification from the Auto Maint tab, you need to import Cal and Ver kit information. Perform that procedure from the Lot Management tab. See Importing CAL or VER Kits. Run the Calibration/Performance Verification routine as part of your weekly maintenance routine: 1. On the Home page, click System Initialization under Daily Activities. The Auto Maint tab on the Maintenance page opens. 2.
Active Reagents - Use this section to change calibration or performance verification kits, lot numbers and expiration dates, or to delete a kit. • Calibration Kit - Select a calibration kit from this list. Details of the selected kit display in the Lot Type, Active Lot Number, and Expiration Date fields. • Performance Verification Kit - Select a performance verification kit from this list. Details of the selected kit display in the Lot Type, Active Lot Number, and Expiration Date fields.
• Verifier - If a verification reagent is selected, the Lot Details table displays target information. • Sheath Fluid - Click Edit, enter the Current Lot# and Expiration date. • Import Kit - Opens the Import Calibration or Performance Kit dialog box to import a kit. • Import - Opens the Import Calibrator or Verification Lot dialog box to import a lot. • Export - Opens the Export Calibrator or Verification Lot dialog box. Select a name and location to save the calibrator or verification lot and click Save.
• Remove Air Bubbles (Luminex) • Remove Clog (Luminex) • Revive After Storage (Luminex) • Verification (Luminex) • Weekly Maintenance (Luminex) Commands - This section contains the following commands: • Wash - Sends distilled water through the fluidic lines in the system. It pulls the fluid from a well or from the reservoir and runs it completely through the system to the waste receptacle. • Soak - Prevents salt crystals from forming in the probe due to air exposure.
NOTE: You should perform a Self Test as part of your scheduled weekly maintenance. Click a command to add it to a new routine, to modify an existing routine, or to run the single command. It will appear in the plate image to the right of the Commands. You can also select a location first, which places the command where you choose. To change the location of a command already on the plate, select it in the Command Sequence, and then click a different well or reservoir in the plate image.
6. Click Save As to save the new routine. The Save Routine dialog box opens. 7. Type the routine name in the Routine Name box, then click OK. Editing a Routine 1. Open the Maintenance page, then open the Cmds & Routines tab. Click the routine that you want to edit from the Routine Name list. 2. Click a command in the Command Sequence list that you want to edit. Click Clear to delete the command, or click in a different well in the plate image to change the location of that command.
2. Click Import. 3. In the Open dialog box, browse for the file you want to import, then click Open. The routine becomes the active routine. Exporting a Routine 1. Open the Maintenance page, then open the Cmds & Routines tab. 2. Click Export. 3. In the Save As dialog box, navigate to the folder where you want to store the routine file, then click Save. NOTE: If you haven't previously saved a routine, the Export button will not display.
• Plate Location pin - A green pin that shows the well location used to auto adjust the probe height for the main plate, reservoir, and/or strip wells. The default (and recommended) location for the 96-well plate is well D6. • The main plate depicts a 96-well microtiter plate. For information about the plates used in assays, see Plates. • Strip Well depicts the strip well added to the off-plate reagent area when extra wells are required.
8. In the Reservoir section, click in an off-plate position. 9. In the Strip-Wells section, click on a well location. 10. Click Auto Adjust Height. The probe automatically adjusts itself to the plate you selected. 11. Click Save plate.
Use this tab to view information and diagnostics about the Luminex instrument.
• System Temperature • DD Temperature • CL1 Temperature • CL2 Temperature • CL3 Temperature • CL1 Voltage • CL2 Voltage • CL3 Voltage • DD Voltage • RP1 Voltage • Number of Syringes Items in this list relating to calibration and verification (CAL/VER) have one of the following states: • Passed - Indicates that the process completed successfully. • Failed - Indicates that the process was not completed successfully. Failed items appear in red. • Not Current - Indicates that verifiers are not current.
• Log Date • Message • Code • Export - Opens the Export System Log dialog box. Select a name and location for the system log export and click OK. Select Overwrite to overwrite a pre-existing file. This button exports the file in .csv format. • Print - Click to print the log file. • Save - Click to open the Save As dialog box. This saves the file as a .pdf. Pick a file name and location to save the file, and click Save.
Support Utility Tab Maintenance > Support Utility Use this tab to type information that will be useful in the event you need to contact Luminex Technical Support. On this tab, you can type your contact information and comments, as well as create and export a .zip file containing system logs and optional batch files. This file can be sent to Luminex Technical Support, who can use the information in the event of a service call. This tab contains the following: • Include Batch Info.
• Clear - Clears all the information in the Name, Phone, Company, Email, and Comment boxes. • Browse - Opens the Browse for Folder dialog box to choose an alternate destination folder for the utility .zip file. Sending a Support.zip File 1. If you want to include a batch file, select it and check Include Batch Information. 2. Press Support. The Support Utility launches. 3. Type your name in the Name box. 4. Type your company name in the Company box. 5. Type your phone number in the Phone box. 6.
Chapter 8: Admin Page You must have the appropriate license to view some of the features of in the Admin page.
Use this tab to configure system settings such as application settings, LIS settings, maintenance page options, and external analysis program settings. Application Settings These settings change some of the basic ways that xPONENT works, allowing customization of many of the basic functions. • Enable Virtual Keyboard - Enables the touchscreen virtual keyboard. • Allow the application to be minimized - Enables minimization of the xPONENT software so that the computer’s desktop can be accessed.
External Analysis Program Settings This option is for those using a program other than xPONENT® to analyze collected data. • Installed Analysis Programs - Lists the analysis programs currently installed. • Add New - Opens the New External Analysis Program dialog box. • Browse - Opens another dialog box to select the file location for the third-party analysis program. The selected location appears in the Path box, and the name displays in the Name box.
• System Initialization Procedure - Displays the available selections to use as the default system initialization procedure. After selecting an option, click Save at the bottom right portion of the window to save the selection as the default procedure. • Allow running calibration or verification if the instrument is not warmed up - You can enable or diable this feature by selecting the check box. • Calibration expiration days - Allows to set how often the system will need to be recalibrated.
NOTE: The Home page, and in some instances the Admin page, cannot be moved. To arrange the main navigation buttons: 1. Select or clear the check boxes by each page name to hide or display the page. 2. Click a page name and use the up and down arrows to change the order in which the pages display, from left to right. 3. Click Save. 4. Click Default if you want to restore the main navigation. Maintenance Options This section allows you to define the System Initialization Routine.
2. Click System Setup. 3. Under Maintenance Options, you can select a procedure from the drop-down list. • Laser warm-up, fluidics prep, calibration, performance verification • Laser warm-up, fluidics prep, performance verification • Warmup, fluidics prep 4. Click Save. Group Setup Tab Admin > Group Setup This tab is accessible only in the Security or 21 CFR Part 11 packages. Use this tab to assign permissions to different groups of users.
Group Profile - The following user groups are predefined: • Administrator • Supervisor • Service • Technician2 • Technician1 • Reviewer The user will belong to the group you select. Group Features - The Group Features list contains permission categories. When you select a category from the list, the Features section displays the individual tasks that are a part of that category.
• Validate and Invalidate Results • Approve Batch • Reanalyze Results • Save Batch after changing results • Change Formula • Reacquire errored wells for partial batch • View Processed Batch Results • Export Processed Batch Results • Change Sample Load Volume During Run Protocol Management Delete Protocol Lot and Std/Ctrl Kit management • Create Std/Ctrl Kit and Lots • Edit Std/Ctrl Kit and Lots • Delete Std/Ctrl Kit and Lots Import and Export Data • Export Batch, Protocol, Kit or Lot Files • Import Batch, P
You must have administrative privileges and be using the Secure version of xPONENT or the 21 CFR Part 11 version to perform the following tasks. • Create New User - Opens the Create User Account screen. • Global User Settings - This section contains the following: • Password Expiration - Type a length of time in days. The default is 180 days. • Unsuccessful Login Attempts - The default number of allowed unsuccessful logins is three attempts.
Restoring Account Status If users attempt to log in unsuccessfully more than the number of times allowed, they will be locked out. In the User list, click the user ID, then click Edit User. Clear the Account status: Locked check box, then click Save. Create User Account Window Admin > User Setup > Create User Account Use this tab to set the options for a new user. Any user with access to this tab can assign rights to any and all groups.
• Technician2 • Technician1 • Reviewer • Password - Type a unique password for the user in this box. • Reenter Password - This becomes active when you type a password in the Password box. Type the password again in the Reenter Password box to confirm it. • Change password after next login - Requires the user to change the password after the first login. This check box is enabled by default. • Save - Saves and returns to User Setup. • Cancel - Returns to User Setup without saving.
• Group Profile - The user's group or category. • Change Password After Next Login - A box that, when checked, indicates that the user will change his or her password at the next login. • Account Status - Locked or functioning. A check in the Locked box indicates that the account is locked due to the maximum number of unsuccessful login attempts. • Password - The user's current password. • Reenter Password - The user's current password, repeated.
• Allow running a batch if the instrument is not warmed up - Allows batches to run if the instrument is not warmed up. • Allow running or saving a batch with expired reagents and fluids - Allows batches to run with expired reagents and fluids. • Allow running a batch if XY temperature is not in range - Allows batches to run if the XY temperature is not in range. • Allow running a batch if the instrument is not calibrated or verified - Allows batches to run if the instrument is not calibrated.
• Error (Stop) - Stops the batch. • Warning ( Pause) - Pauses the batch. • Warning (Log) - Logs a warning if the number of successive wells in the Well Count box are run without reaching the number of beads in the Bead Count box. • Run Routine - Displays a drop down box which allows you to select the routine you want to run when a low bead count is detected. Click in either box to type a number other than the default.
• Scheduled Maintenance - A scheduled maintenance routine is now due. • Low Bead Count Detected - A bead count below the accepted range is encountered. • System Alerts - One of the following events has occurred: • Overloaded syringe pump plunger • Probe step loss • Sheath pressure error • X step loss • Y step loss • Routine Complete- The currently running routine is complete.
4. If you choose to send an email notification for a specific event, and the email address is not already set up, click Setup Email to open the Setup Email dialog box. • Turn email notifications on or off by checking or clearing the Email Active check box. • Contact your system administrator to determine the correct information for the Mail Server Host, From Email Address, From Email Password, and Mail Server Port boxes, and if you need to check the Enable SSL (Secure Sockets Layer) check box.
CSV and Batch Export Options This tab contains the following: • Automatically export results CSV file when batch is complete - Automatically export the results .CSV file when the system finishes analyzing the batch. This allows running programs on exported data without having to manually start the export. • Automatically export batch when batch is complete - Exports batch information automatically when a batch has completed.
• Automatically convert the raw run files to CSV format for each well in the batch Click the check box if you want the automatic conversion. • Cancel - Cancels changes. • Save - Saves changes. Archive Options Tab Admin > Archive Use this tab to archive all file types - batches, protocols, kits, lots, and LIS records. When you click the Launch Archive Utility button, the Archive Utility opens. NOTE: To back up or restore xPONENT files, quit the application and select XPONENT ARCHIVE.
The Archive Utility can be used within xPONENT to set up frequency and type of files to back up. Statistics about disk space and archive space used, as well as the types of files to backup, appear on this page. The archive utility runs continuously. If automatic archives are scheduled, they occur whether xPONENT® is running or not. When you want to backup or restore xPONENT, quit the application and run the Archive Utility from the Start menu.
• System Restore - Click this to restore your system. It requires a log in and then refers you to the Start menu to restore your system. • Manual Archive - Click this to perform a manual archive process. It requires a log in and then opens the Manual Archive window. • Close - Click this to close the Archive Utility.
Setting Up Automatic Archiving Click Edit in the Schedule Overview box to enable the fields to accept your edits and to enable the Browse button. The label on the button changes to Save. 1. Specify when you want your archives to begin using the calendar. 2. Using the drop-down list, select how often you want to perform archives. 3. Select Reminder Only to be reminded of the need to archive, after which you must archive manually. Clear this check box to enable automatic archives. 4.
1. In the Archive Utility, click Manual Archive to open the Manual Archive window. 2. Using the tabs on the left side of the window, select the type of files you want to archive: • Batches • Logs • Protocols • Kits • Lots • LIS Records Each tab opens a list of files available for archiving, except Logs, which requires only a choice of how old, in days, a file has to be to be archived. 3.
Contact Luminex Technical Support to upgrade xPONENT® for FLEXMAP 3D® or to obtain a new license. NOTE: You must restart the computer for the new license to take effect. Adding a New License Key 1. Access the Admin page, then the Licensing tab. 2. Click License (bottom right corner of window). 3. Copy and paste the new key into the License Code field. The License File field remains blank. 4. Click OK. This closes xPONENT®, applies the license, and restarts xPONENT®.
Use the Schedule tab to enable or change the recurrence of scheduled maintenance reminders. • Select the time of day that you want to receive alerts in the Alert Time list. • Select how often you want to receive reminders in the Recurrence list. • Enable or disable the reminders by selecting or clearing the Enabled check box. Editing Maintenance Schedule Settings Edit maintenance settings on the Schedule tab accessed from the Admin page.
2. On the Schedule tab, use the drop-down menus to edit the following items for any scheduled activity: • Alert Time - the time of day that you want to receive alerts. • Recurrence - how often you want to receive reminders. • Laser Warm Up Schedule - you can schedule what time you want the lasers to warm up. You can either enable or disable this option. 3. Enable or disable the reminders by selecting or clearing the Enabled check box.
1. Type a company name in the Company box, and any additional information in the Info: box. NOTE: The logo file should be 920 x 125 pixels. If you want the logo to appear to the right of your company name, include 120 pixels of white space to the left of the logo, in the graphic file. If you do not include white space, the logo may appear behind the company information. 2. Click Import Logo to open the Open dialog box and select the file you want to use for the logo at the top of reports. 3.
