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Muse™ Cell Analyzer

1. Count amp Viabilit Annexin V amp Dead Cell empty empty n Select a favorites item to edit on the Home Page User ADM Status Bor jons Select the assay that you want to add or replace for the selected slot Or select Remove Item to remove the assay from the selected slot Home gt Setup gt Edit Favorites Re move tem Edit Item All Assays apeina Dead gal amp Viabil Select an Assay to insert or Select Remove Item User KG Status Bor fy Setup 79 The Edit Favorites screen now appears with the selected assay in the slot or the assay removed from the slot 5 Select Save Managing Instrument Data Storage The Manage Instrument Storage options allow you to import data to and export data from the system Use the export features to back up or share data For a description of the folders created on the USB drive when you export data see Folder and File Structure on page 19 1 Select Setup from the main menu 2 Select Manage Instrument Storage from the Setup menu 3 Select the option you wish to perform Only administrators can import and export all user data Operators can export their own data by user and date Maximum available storage 160GB Current storage usage 45GB Displays the free disc ee and how much space is being used Export by User Export by Date gt Import
2. STATUS STATUS 72 ipn 0040 44 40 44 CTATI IC 4 Waste Cy A STATUS STATUS STATUS End of Cleaning End of CI Waste Cy End of W Cleaning End of CI End of W Cleaning End of CI End of W Cleaning USB1 My 4GB v 201 1Dec08_141423 log Ld Select the appropriate check box es to display the types of events you wish to view Filter the event log list by selecting the types of events you wish to view Optional If you wish to export the cleaning event log select Export from the Event Log screen select the USB drive from the Export Location field enter a file name or use the default name and select Export 62 Muse Cell Analyzer User s Guide Resetting Fluid Levels Use the Reset Fluid Levels option to reset the status indicator for the waste bottle and cleaning solution bottle back to 100 and 100 after you have emptied the waste bottle and filled the cleaning solution bottle It is important to reset the fluid levels each time you fill and empty the bottles so that the instrument can accurately determine the amount of fluid in each bottle capacity remaining fluid remaining User ADM Status Cleaning Complete f 1o hn 100 When the waste bottle has 1 capacity remaining or the cleaning solution bottle has 1 fluid remaining a message appears prompting you to fill the cleaning solution bottle and empty the waste bottle 1 Select Clean from the System Ch
3. An a lyze r AVOID DIRECT EYE EXPOSURE TO BEAM EMD Millipore Corporation 25801 Industrial Blvd Hayward CA 94545 510 576 1400 15 V 5A Complies with 21 CFR ns 1 Chapter 1 Subchapter J li A in USA with Aa of Foreign Origin C amp s c U The M logo is a registered trademark of Merck KGaA Darmstadt Germany Muse is a trademark of Merck SN 7200120001 4000 5880 Rev A KGaA Darmstadt Germany Limitations e The Muse System is for research use only not for use in diagnostic or therapeutic procedures e The results of the assays are dependent upon the proper use of reagents and instrument Refer to the appropriate kit user s guide located at www millipore com muse for specific instructions and limitations 4 Muse Cell Analyzer User s Guide Introduction Muse System The Muse System consists of a compact portable and easy to use cell analyzer software and optimized reagents The Muse Cell Analyzer uses patent pending miniaturized fluorescent detection and micro capillary technology to deliver quantitative cell analysis of both suspension and adherent cells 2 to 60 um in diameter The software includes a dedicated module for each assay as well as a tools for verifying instrument performance and cleaning the instrument s fluid system Convenient mix and read assays are optimized for accuracy and convenience The available assays include e Count amp V
4. Muse Cell Analyzer User s Guide For Research Use Only not for use in diagnostic procedures Copyright Disclaimer Limitations Trademarks Patents 2012 Millipore Corporation Inc All rights reserved No part of this publication may be reproduced transmitted transcribed stored in retrieval systems or translated into any form or by any means electronic mechanical magnetic optical or otherwise without the prior written permission of Millipore Corporation 25801 Industrial Blvd Hayward CA 94545 United States of America Millipore reserves the right to change its products and services at any time to incorporate the latest technological developments This guide is subject to change without notice Millipore has not validated the analysis of Muse System data using third party programs and cannot warrant that the results using these programs will be correct Millipore does not provide support for any third party programs The Muse System is for research use only not for use in diagnostic or therapeutic procedures The results of the assays are dependent upon the proper use of the reagents and instrument Please refer to the appropriate reagent kit user s guide for specific instructions and limitations The M logo is a trademark of Merck KGaA Darmstadt Germany Millipore Muse and guava are registered trademarks or trademarks of Millipore Corporation Microsoft and Excel are registered tradem
5. 6 Select Close to the message that data was successfully exported Software Update Use Software Update to load the latest version of Muse software on your instrument 1 Download the software update from www millipore com onto a USB thumb drive 2 Install the USB drive into an available port on the instrument Select Setup from the main menu 4 Select Software Update from the Setup menu The system automatically scans the available USB drives for a software update file If an update is found a confirmation message appears Setup 85 5 Select YES to install the software update Software Update detected This will update the software from version 1 0 to version 2 0 Example Are you sure you want to continue Cancel The new software will install and the system will automatically reboot in the new version A message appears that the software successfully installed The Select User screen appears Service Tools Service Tools allows a qualified service technician to connect a laptop and run a service application for troubleshooting purposes 86 Muse Cell Analyzer User s Guide Troubleshooting This section lists possible problems you might encounter or messages you might see during operation If you have a problem that you cannot resolve by using the troubleshooting section call technical support toll free in the USA and Canada at 1 800 MILLIPORE 645 5476 or visit www millipore com techser
6. ADU Status Ready for System Check gu t 42 Muse Cell Analyzer User s Guide Printing Results To print the results select the print tab to the right of the Bead Info tab on the Results screen You can choose to print the current results the log list see Viewing Past System Check Results on page 44 or a trend graph 1 Select the printer from the drop down list of printers The list displays all available printers If no printers are detected the default printer is PDF to USB which allows you to export a PDF 2 Select the Print Current Check button to print the current results You can also print the entire log list past System Check results or the trend graph If you select to print the trend graph the number of runs included in the printout is based on the number of runs selected at the Trend Graph screen see Viewing a Trend Graph on page 47 3 Optional Select Print Preview if you want to view the results as they will print 4 Select Print If no printers are available or if you wish to export to a PDF select PDF to USB then select Print The Export to PDF screen appears Select the USB port enter a file name for the report file or leave the default name Select Export 11 0254 07 SEP 2011 07 45 23 v a Log List button es Print tab Printer HP 675 All in One v Status Ready Paper Size 8 x 11 a Print Current Check Print Preview C Print Log
7. embedded Intel ATOM based computer 160 GB data storage output data file formats binary data storage in Flow Cytometry Standard FCS 3 0 format spreadsheet results file in comma separated value CSV format 10 lt 10 CV Following is a list of supported printers e HP Officejet 100 Mobile Printer e HP Officejet 6000 Wireless Printer e HP Officejet Pro 8000 Printer Series A809 98 Muse Cell Analyzer User s Guide Compliance The Muse System contains a Class Illb laser operating at 532 nm This product complies with e CFR Code of Federal Regulations Chapter 1 Subchapter J and installation overvoltage category II e Class limits for exposure to laser radiation set by the Center for Devices and Radiologic Health CDRH Symbols Symbol Meaning Attention consult accompanying documents gt C Affixed in accordance with European Council Directive 73 23 EEC Danger laser radiation P In accordance with Canadian Standards Association amp O c 7 Separate collection of waste at end of life as required by European Directives Dispose of in accordance with the applicable country regulation gt px Dangerous voltage Specifications 99 Warranty EMD Millipore Corporation Millipore warrants its products will meet their applicable published specifications when used in accordance with their applicable instructions for a period o
8. and is concentrated in an intense narrow beam A boundary or set of boundaries used to segregate data into subsets for statistical analysis Set a marker on a histogram to obtain statistics on a certain region Set quadrant markers on a dot plot to obtain statistics on data within four quadrants The average of the fluorescence intensities of each event acquired within a given set of events 92 Muse Cell Analyzer User s Guide median The axis value for the event that falls in the middle of the distribution parameter A specific cell property that is measured as the cell passes in front of the laser beam Each parameter is the output from a photomultiplier which measures fluorescence or a photodiode which measures forward scatter population A group of cells that express similar values within one or more parameters For example cells that are positive for a particular antibody appear in the same location within a histogram or dot plot red detector A device used for measuring light intensity The red channel on the Muse Cell Analyzer can measure fluorescent light from dyes that emit signals similar to PE Cy5 threshold The minimum level of discrimination to electronically eliminate unwanted signal A threshold setting allows you to specify events you wish to acquire based on signal intensity of the event Anything below the threshold in not acquired yellow detector A device used for measuring light intensity The yel
9. any assay screen Touch the title bar to open the About dialog Muse Version 1 0 0 0 Instrument ne 20001 1 Firmware V0 OS 1 0 0 0 Copyright 2012 by EMD Millipore 25801 Industrial Blvd Hayward CA 94545 www millipore com muse Technical Support 1 800 MILLIPORE 1 800 645 5476 www millipore com company sup3 support EMD Millipore is a division of Merck KGaA Darmstadt Germany Close Close Window 26 Muse Cell Analyzer User s Guide Getting Started To prepare the system for use perform the following steps each day that you use the instrument e Empty the waste bottle and add bleach See Emptying the Waste Bottle on page 65 e Fill the cleaning solution bottle with ICF See Filling the Cleaning Solution Bottle on page 64 e Reset the fluid levels See Resetting Fluid Levels on page 63 e Perform a Complete System Clean See Performing a Complete System Clean on page 55 Turning On the Unit 1 Ensure the instrument is plugged in BI NOTE If you are plugging in the instrument for the first time it will automatically turn on After a brief startup sequence the logon screen appears Refer to Logging On as the First User on page 29 2 Press the power button located on the front of the unit below the touch screen A screen appears allowing you to select your user name from a list Refer to Logging On Once Users are Added on page 32 Getting Star
10. are entered Refer to the System Check Beads vial label and information card for values 2 Run Quick Clean then rerun the System Check procedure If results still fail run the Complete System Clean procedure 3 Prepare a fresh bead sample and rerun the System Check procedure 4 Rerun the Complete System Clean procedure Ensure that the cleaning solution vial has sufficient fluid and that there are no kinks in the tubing from the flow cell or the cleaning solution vial Data sets saved by the default Administrator user are no longer accessible after a new Administrator user is added to the system The default Administrator is overwritten when a new Administrator user is added to the system Add another administrator level user with the name Administrator Sample acquisition is taking longer than expected or progress bar stops during acquisition Ensure that the System Check procedure was run and passed If the progress bar stops during acquisition the fluid system may be clogged Run Quick Clean Instrument clogging too many cells Run Quick Clean to clean out the capillary Resume running sample Lower Cell Concentration warning during acquisition The sample concentration may be too low The assay instructions are optimized to give you cell concentration between 100 and 500 cells L for accurate results Repeat sample preparation with a lower dilution factor to allow for adequate cell numbers Hig
11. are intended for research use only and are not to be used for any other purpose which includes but is not limited to unauthorized commercial uses in vitro diagnostic uses ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals 100 Muse Cell Analyzer User s Guide EMD MILLIPORE For Ordering Information or Technical Assistance In the U S and Canada phone 1 800 645 5476 fax 1 800 645 4539 www millipore com techservice For other countries across Europe and the world visit www millipore com offices Millipore Corporation 25801 Industrial Blvd Hayward CA 94545 USA 2012 EMD Millipore Corporation All rights reserved 0110 7895 Rev B Printed in U S A April 2012
12. bottle flow cell access door touch screen tube holder sample loader arm power button System Overview 9 Sample Loader The sample loader holds an individual sample tube The loader arm can be lifted easily with your finger and released with the touch of a button using the touch screen The following tube is supported e 1 5 mL microcentrifuge tube with conical tip and screw cap if snap cap tubes are used cut off the cap The system automatically detects tubes with caps and will not load the tube if you install a tube with a cap Loading and Unloading a Tube To load a tube place the tube in the tube holder and lift up on the loader arm As you lift the arm the tube automatically slides back to the loaded position and is seated under the capillary During a run the loader arm drops automatically when sample acquisition is complete To manually unload a tube use the eject button on the touch screen Additionally a lever located in the flow cell access door allows you to drop the loader arm to remove a sample tube in the rare event of a power outage Open the flow cell access door and push back on the switch shown in the following illustration loader arm release lever Push back to drop loader arm 10 Muse Cell Analyzer User s Guide Cleaning Solution and Waste Bottles The cleaning solution bottle can be filled with Guava Instrument Cleaning F
13. main menu 2 Select Users amp Access Levels from the Setup menu 3 Select Add New User M NOTE All fields are required 4 Enter the user name using the keypad 74 Muse Cell Analyzer User s Guide Enter up to 3 characters in the Initials field to identify the user Select Done These initials will appear in the status bar when the user is logged on Enter up to 4 numeric characters for the password Select Done Confirm the password by reentering it Select Done Select an access level Administrator or Operator Select Add User Name Initials Katie Gillis 9 ko 92 Password Confirm Password Access Level Administrator Operator Cancel Add Cancel Add Addr er User ADM Status 100 yes Removing a User Only administrators can remove a user from the system The default Administrator will be overwritten by the first Administrator level user added to the system The default Guest user cannot be removed A Cc N NOTE Removing a user does not remove the user s FCS data from the system Select Setup from the main menu Select Users amp Access Levels from the Setup menu Select the user name from the list of users then select Remove User Select Yes at the confirmation screen to remove the user Setup 75 Editing User Information Any user can edit their user name and or password Only administrators can change a user s access level 1 2 3 4 Select Setup
14. o See 24 Muse Cell Analyzer User s Guide Data Set Options Use the Options button on the Results screen to access features that allow you to rename the data set export data save instrument settings and view the event log 1 Select Options in the title bar of the results screen The Data Set Options screen appears Rename Data Set Export Data Set Export to Spreadsheet Save Current Settings Event Log l Close Close Window e To rename a data set select Rename Data Set edit the file name and select Apply You can also rename the data set at the completion of the run when you select Finish e To export the data set select Export Data Set select the USB drive from the Export Location field enter a file name or leave the default Select Export You may want to export the data set so that you can share the data with other users e To export the data to a spreadsheet file select Export to Spreadsheet select the USB drive from the Export Location field enter a file name or leave the default Select Export e To save the current instrument settings see Saving Instrument Settings on page 23 e To view or export the event log see Event Log on page 17 System Overview 25 About Dialog The About dialog provides the software version number and copyright information as well as technical support information To access the About dialog touch the title bar of the main menu or
15. screen in preparation to run the first sample e Select YES if you wish to perform the adjust settings step using the settings you just retrieved This will help to ensure that the retrieved settings are correct Saving Instrument Settings 1 2 3 Select Options from the Results screen after the sample is acquired Select Save Current Settings Select the location where you want to save the file The default directory is the MUSE directory on the unit s hard drive If you want to save the file to a USB drive select the USB drive from the Save In field A new directory will be created on the USB drive MUSE user namelassay name instrument settings file Enter a file name or use the default file name Select Save MUSE v PT_30MAR2012_140027 Cancel Cancel Save System Overview 23 Assay Overview Detailed steps for each assay are covered in the reagent kit user s guide which can be found at www millipore com muse Some additional information on various tasks that can be performed from the assay screens is listed in the following section Home Assay Settings Run STU E Options 002Sample 002 4 Sample Info Viable Cells mL 2 68E406 Viability 60 30 Example of Count amp Viability Total Cells mL 4 45E4 06 results screen Select plot to adjust marker gate POPULATION PROF ILE 4 VIABILITY PROFILE a 8 E a CELL SIZE INDEX TE User KG Data Set KG 14D CZUT 144947
16. the results for an individual sample as well as all acquisition information at the time of data collection FCS files are defined by the Data Files Standards Committee of the Society for Analytical Cytology Cytometry 1990 11 323 332 Glossary 91 flow cell fluorescence fluorochrome gate histogram laser marker mean fluorescence An optical assembly within the Muse Cell Analyzer The flow cell consists of a metal shuttle holding a glass capillary with a tiny chamber where the laser beam illuminates the sample stream and cellular measurements occur The phenomenon of light emission that occurs when a fluorochrome s excited electrons drop to a lower energy level A fluorescent dye used as a detection reagent in cell analysis applications A molecule capable of absorbing light at a certain wavelength then emitting light at a longer wavelength fluorescence as it releases energy A graphical boundary that defines a subset of data Gates may be set on a single parameter histogram or a two parameter dot plot A graphical representation of single parameter data The horizontal axis of the graph represents the increasing signal intensity of the parameter and the vertical axis represents the number of events cells Light amplification by stimulated emission of radiation A light source that is highly directional monochromatic coherent and bright The emitted light is in one or more narrow spectral bands
17. the main menu 2 Select Manage Instrument Storage from the Setup menu 3 Select Export by Date to export data for a particular time frame 4 Select the USB drive from the Export Location field 5 Select the files to export 30 60 or 90 days or older 6 Ifyou wish to delete the data once it has been successfully exported select Delete files from instrument after successful export M NOTE Only administrators can delete data after exporting 7 Select Export If you selected to delete the data after exporting a dialog appears requesting confirmation Select Yes Otherwise a message appears indicating the export status Select Close Delete files from instrument after successful export Export Location E USB1 My 4GB v Select files to export w 30 days or older ka 60 days or older Ww 90 days or older Back Export peer statu Bs fy 82 Muse Cell Analyzer User s Guide Importing All User Data If you exported all user data for example if you sent your system out for depot service you may wish to import the data back to the system Only administrators can import all user data A WARNING Importing all user data will overwrite any data that you currently have on your system 1 Select Setup from the main menu 2 Select Manage Instrument Storage from the Setup menu 3 Select Import All User Data 4 Selectthe USB drive where the data is stored If you wish to also import the data in the Public folder select Include Public Folder 5 Se
18. this folder structure O Muse O User Name O Assay Name Data set files fcs csv Spreadsheet files csv Event log log Instrument settings files gst Assay reports pdf O System Clean Cleaning log log Cleaning log pdf O System Check Service Check file zip O Replicate1 CHK fcs O Replicate2 CHK fcs O Replicate3 CHK fcs O MuseCheckLog csv System Check log csv Event log log E Trend Graph report pdf E System Check report pdf O Public Oo e o Files folders generated by Muse software Files generated by users through Export Files generated by users through Print to PDF function The Public folder is created automatically and allows administrators to share data If an administrator wants other users to have access to certain data and or instrument settings files they can export the data to the Public folder While operator level users can access this data they cannot modify the files or save files to the Public folder Bi NOTE The System Check and System Clean folders are created only if a user exports System Check and System Clean files System Overview 19 File Storage The Muse System has an internal hard drive for saving data files To see the available storage select Setup from the main menu then select Manage Instrument Storage The amount of available storage is dis
19. use System Overview 11 Software Overview Use the software to acquire samples view results and control the instrument In addition each software module allows you to adjust the markers and or gates to fine tune the analysis Instructions for running samples using the individual assay modules can be found in the reagent kit user s guide for each individual assay kit Navigating Through the Software A navigation bar at the top of the screen indicates where you are in a given procedure The current step is highlighted gray These buttons vary depending on the procedure you are performing for example an assay Complete System Clean or System Check procedure If you access one of these procedures and wish to exit simply press Home to return to the main menu If the screen you re on does not display a navigation bar select the Back Close or Cancel button from the current screen to access a screen with a navigation bar For simplicity the navigation bar has been cropped out of most of the screens appearing in this guide Home hec Info Run Results Navigation bar A status bar at the bottom of the screen indicates the user currently logged on instrument status and the volume remaining in the cleaning bottle and capacity remaining in the waste bottle User KVT Status Ready for Check il 23 20 Example shows waste bottle has 23 capacity remaining 77 full and cleaning bottle has 20 of the solution remainin
20. 0 data file and a CSV spreadsheet results file each containing data for all samples in the run Additionally you can optionally choose to save a separate file of the instrument and analysis settings FCS 3 0 Data Files A single FCS file includes the results and sample information for all samples acquired within a run as well as an event log At the completion of the run after you click Finish to save and close the data set you can enter a new file name or use the default file name that appears The default file name is the user s initials followed by the date and time for example PT 15Feb2012 115618 FCS The file name assigned to the FCS file is also assigned to the spreadsheet results file An extension is automatically appended to the file name you enter The first three characters represent the assay type followed by FCS Assay File name Count amp Viability filename VIA FCS Annexin V amp Dead Cell filename NEX FCS Cell Cycle filename CCY FCS Spreadsheet File A spreadsheet file containing the data for all samples within a data set is automatically saved along with the FCS file in the same directory The spreadsheet file contains a summary of the statistical results sample information and instrument settings for each sample run within a data set It can be opened and analyzed using a spreadsheet program such as Microsoft Excel The same file name assigned to the FCS data file is also used for th
21. All User Data gt Only administrators can perform Export All User Data gt these functions Back 4 Refer to the following sections for more information 80 Muse Cell Analyzer User s Guide Exporting by User You can export data by the user Operator level users can only export their own user data 1 2 3 4 5 Select Setup from the main menu Select Manage Instrument Storage from the Setup menu Select Export by User to export data for a particular user Select the USB drive from the Export Location field Select the user whose data you wish to export from the Users list Use the arrow buttons in the bottom right corner to scroll through the pages of users If you wish to delete the data once it has been successfully exported select Delete files from instrument after successful export BI NOTE Only administrators can delete data after exporting Select Export If you selected to delete the data after exporting a dialog appears requesting confirmation Select Yes Otherwise a message appears indicating the export status Select Close Delete files from instrument after successful export Export Location USB EJ v Users Julie Clor WP Katie Gillis e Guest 4 iol P User JC Status fe fy Setup 81 Exporting by Date You can export data from the last 30 60 or 90 days or older Operator level users can only export their own user data 1 Select Setup from
22. For this reason it is a good idea to save your PIN in a secure place If you are the sole administrator and you forget your PIN you can export your user information to a USB drive and send it to Millipore We will send a PIN back to you which you can then import Getting Started 33 1 If you forgot your PIN select Done or touch outside the keypad to close it then select Forgot User PIN Enter User PIN Username Katie Gillis g Remember User PIN Forgot User PIN Cancel 2 Insert a USB drive 34 Muse Cell Analyzer User s Guide 3 Select Export User Info to USB thumb drive M NOTE Be aware that once you export your PIN you will no longer be able to log onto the system even if you should remember your PIN Enter User PIN Username Katie Gillis g Remember User PIN Export User Info to USB thumb drive gt Use PIN from Zip in USB thumb drive gt 4 Select the drive you want to export to then select Save Contact Tech Support You will be asked to e mail the zip file from your thumb drive to Tech Support 6 Tech Support will e mail you another file which you will save to a USB drive 7 insert the USB with the file from Tech Support Begin to log on select your user name and Forgot User PIN Then select Use PIN from Zip in USB thumb drive 8 Follow the steps to reset the PIN Getting Started 35 Selecting an Assay to Run The main menu displays the following assays e Count a
23. I water Sample Loader Select Continue ar Ur Os When the system cleaning procedure is complete the Cleaning Log appears Each entry in the log represents one System Cleaning procedure The list shows the user who performed the cleaning the date and time the cleaning was performed and whether the cleaning was completed or aborted 26 JAN 2012 14 40 57 IA Log List Scroll through cleaning log entries User 25 JAN 2012 13 03 43 Complete 19 JAN 2012 03 07 PM Complete Finish iom User ADM Status Clesring Complete Be LES Cleaning and Maintenance 57 If you are shutting down the system leave the tube of DI water used for cleaning loaded on the unit and select Finish to return to the main menu A WARNING Do not leave a tube of Guava ICF bleach or any other cleaning agent loaded on the instrument overnight or for an extended period of time Prolonged exposure to strong oxidizing agents will damage the flow cell Always leave a tube of DI water on the system after cleaning and when shutting down Change the tube of water regularly to ensure it is clean and free of particles Optional To run the cleaning procedure again select Next Cleaning Printing the Cleaning Log You can print a list of all Complete System Clean procedures performed since the last time the cleaning log was cleared The list contains the date and time that each Complete System Clean was run as well as the operator
24. OF Print Trend Graph Finish EE Next Run Return to Launcher Run Next Check Print results log of all results or trend graph User KVT Status Perlormence Check Completed ij 25505 System Check 43 Viewing Past System Check Results To see the results from previous runs select the Log List button below the Clean button on the Results screen You can also use the arrows to the right of the Log List button to quickly scroll through and display previous System Check results in the Bead Info tab Each entry in the System Check Log represents one System Check run and includes the user who ran the test the bead lot number the date and time the test was run and whether or not the test passed Y or N A hyphen indicates that the run was aborted Use the arrow buttons at the bottom of the screen to scroll through the pages of results Select a run from the list to display the selected results in the Bead Info tab User Lot No Date amp Time 110279 26 JAN 2012 13 16 13 110279 25 JAN 2012 12 18 18 110279 19 JAN 2012 15 03 16 110279 12 JAN 2012 16 07 01 110279 04 JAN 2012 16 06 48 110279 04 JAN 2012 15 58 48 110279 29 DEC 2011 16 07 47 110279 29 DEC 2011 14 55 55 110279 29 DEC 2011 11 54 20 110279 07 DEC 2011 16 29 52 Cancel Cancel Selection For information on clearing and exporting the System Check Log or viewing a trend graph or the event log see System Check Opti
25. Refer to Users amp Access Levels on page 73 for information on the options available for each user access level Setup Users amp Access Levels Recommended Cleaning Options gt L Printing Layout Options gt Edit Favorites gt Manage Instrument Storage gt Software Update gt Service Tools gt Back User ADM Status is Eject USB D Eject USB drive and adjust volume features available to all users m Setup options Setup 71 3 Select Eject USB in the title bar to safely eject a USB thumb drive The USB drives currently connected to the instrument are listed Select the eject button to the right of the drive you want to eject PERO SST Safely Eject USB unicmcm USB1 My 4GB eject button 4 Select the volume control in the title bar to adjust the audio feedback on the instrument Use the center slider or the arrows on either end of the slider to increase or decrease the volume Select Mute if you wish to turn off the volume Volume Control 4 P Tap arrows Mute Or drag slider Back 5 Refer to the following sections for more information on the Setup task you wish to perform 72 Muse Cell Analyzer User s Guide Users amp Access Levels The Muse System supports two levels of access control Administrator and Operator The access level is assigned by the laboratory administrator when a new user is added to the sys
26. STATLIS Fetahli Example of Count amp Viability event log Close Close Window If errors or warnings occur during a run a message appears in red below the plots indicating that errors warnings have been logged Errors System Check e System Check failed due to avg MFI for channel actual x Target value y System Check failed due to percent CV for channel value x System Overview 17 Warnings System Check Loader arm is down Concentration failed for replicate 1 2 3 Avg conc failed CV failed All assays Sample Loader is down Please raise the Loader arm Adjust Settings timed out Please re enter Adjust Settings if necessary to complete the instrument set up not applicable for System Check The run timed out before enough events were acquired Annexin V and Count amp Viability More than 500 particles uL Sample is too concentrated Please dilute or accuracy may be compromised Less than 10 particles uL Sample is too dilute Accuracy may be compromised Cell Cycle More than 1200 particles uL Sample is too concentrated Please dilute or accuracy may be compromised Less than 25 particles uL Sample is too dilute Accuracy may be compromised 18 Muse Cell Analyzer User s Guide Folder and File Structure When you export data to a USB drive using any of the export features a folder structure is created on the USB drive The following diagram outlines
27. arks of Microsoft Corporation in the United States or other countries U S patents 5 798 222 6 816 257 6 710 871 and foreign equivalents owned by Millipore Corporation Patent App No 61482946 owned by Millipore Corporation The Muse Count amp Viability Reagent is the subject of U S Patent 6 403 378 owned by Millipore Corporation Contents Preface tice aici eaten oe tela eto brad bleus 1 About this Guide diwan eange ni EEEak kaaa p iak 2 Conventions Used in This Guide nananana naana 2 Joc ER 2 Safety vods e eerte tated Bh tae ang Rag Seay epit M pid 3 General Safety i i ce Pad cia eee haga ke d hr 3 Biological Safety 2s Sea ehh ane GER o 3 Electrical Safety zi iss tie eee ete ee DE Eb 3 baser Safety iu ese E Re m be ek urs 3 Precaution Labels 0 0c cece e cece eee eee eee 4 lufrittations c EE A A tant caua naa eae SOS 4 Introductiom sese IG ae uec ee mee exo doe s 5 M se System zie sexus beh eva we wed pork ge Dae ados 5 Setting Up Your System issor cece eee eee eens 6 Components colt dp OU EU YU Ge ae ous 6 Unpacking the Instrument 000 cece eee eee eee 6 System OVEWVieW tace genein greet eeaeen ke 9 Muse Instrument Overview 000 cece cece eens 9 Sample Loader 3x sre REESE ce pias 10 Cleaning Solution and Waste Bottles Luuue 11 Fluid System r crt ERE LER ORTAM 11 Software Overview ssssslslsssssess ee eens 12 Navigating Through the Softwar
28. avorites Count amp Viability Annexin V amp Dead Cell Cell Cycle Essential Tools The following tools are available e System Check to check the system s counting and fluorescence performance prior to running samples The System Check Beads are standard particles used with this tool For more information on running System Check see page 37 e Complete System Clean to clean the fluid system For information on this cleaning procedure see Performing a Complete System Clean on page 55 e Power Options to turn off the system or log off as the current user Essential Tools System Complete Power Check System Clean Options 14 Muse Cell Analyzer User s Guide Entering and Selecting Information The touch screen provides various ways of entering and selecting information most of which are intuitive If you connect an external mouse you can use the cursor to make selections e Touch buttons such as the assay buttons on the main menu e Drag sliders to increase or decrease values or drag handles to adjust gates and markers e Use the keypad to enter information into text or numeric fields Touch the text field to automatically open the keypad Type the information and select Done or touch outside the keypad to close it w Operator Cancel Add CancelAdd User KG Status o fes System Overview 15 Files For each assay the software saves two files an FCS 3
29. cell and replace it with a new flow cell assembly 7 Install the new flow cell The flow cell fits only one way into the receptacle Align the cut out on one corner of the flow cell with the cut out in the receptacle Avoid bumping the capillary tube against the instrument or sides of the receptacle as you install it 8 Close the clamp to secure the flow cell 68 Muse Cell Analyzer User s Guide 9 Reconnect the tubing to the instrument Make sure the tubing is screwed on tightly and there are no constricting kinks or twists 10 Close the access door 11 To ensure that the flow cell was correctly installed run Quick Clean to prime the system Perform a System Check procedure Preparing the Unit for Depot Service Contact EMD Millipore Technical Service for the Decontamination Form and instructions The form must be returned via e mail before you can return the instrument A return authorization RMA number will be issued to you Write this number on the outside of the shipping box If you did not save your original shipping box EMD Millipore can send one to you 1 Perform a Complete System Clean then power off the system 2 Empty and rinse out the waste and cleaning solution bottles and place them in their original box 3 Wipe the outside of the instrument with 70 isopropyl alcohol or a 10 bleach solution in DI water followed by water 4 Open the flow cell access door and disconnect the fluid tubing from the instru
30. ck System Clean Options 28 Muse Cell Analyzer User s Guide 4 Select Power Off to turn off the system Log Off Logging Onto the System The system allows two levels of users administrator and operator For information on the different user levels and access privileges see Users amp Access Levels on page 73 Logging On as the First User The first time you log onto the system you will be prompted to select Administrator or Guest If you select Administrator you will have administrator privileges If you select Guest you will have operator privileges 1 Toadd new users and customize the system select Administrator Bi NOTE Select Guest if you do not have a user account but want to use the instrument Files will be saved with GST initials IMPORTANT NOTE Once you log on as an administrator it is a good idea to add yourself as a user with administrator level access When you add a new administrator level user the default Administrator user shown below will be overwritten For this reason we do not recommended running assays and acquiring data Getting Started 29 sets as a default Administrator as the data will no longer be accessible Muse wv Select a User Default Administrator and Guest users NEN 2 The main menu appears Touch the date and time located in the lower right corner of the screen Essential Tools ij 08 MAR 2012 12 29 Set date and ti
31. e cee e neers 12 Main Menu sre kek jee sit LED SE ICE 13 Entering and Selecting Information 0eeeeee anes 15 I tats certe enipe tia ea a R e d ls 16 Instrument Settings 0 eee eee 21 Assay OVEIVIEW i enne er ea hare e c eed Fue 24 Getting Started sor ee ee ee ee eee 27 Turning Orithe Unit iex eed plod eed vee awe a td es 27 Turning Off the Unit 00 cee eee eee 28 Logging Onto the System 0 cece eee eee 29 Logging On as the First User 0 eee 29 Logging On Once Users are Added 0 cece eee eee 32 Selecting an Assay to RUN 1 1 cece eee eee ens 36 System Check aac d uei veces ia cei EE DIEN NM IM 37 Running a System Check Procedure lsssesssessuees 37 System Check Results 00 0000 e eee eee eens 42 Printing Results osse bee eer eee nen eed aes 43 Viewing Past System Check Results 22 22 000e 44 System Check Options 0 nanan e eee eee 45 Clearing the System Check Log 0 cee eee eee eee 45 Exporting the Check Log 2 eee eee ee eee 46 Exporting the Service Check File cece cece eee 46 Viewing a Trend Graph 0 eee eee eee 47 Viewing the System Check Event Log 00 esses 48 Cleaning and Maintenance leeren 51 Cleaning Protocols seimi aiai ioe cece eee eens 51 Cleaning the Outside of the Unit 0 20 c cece 52 Rinsing the Capillary 0 cec
32. e eects 53 Running Quick Clean 0 cece eects 53 Backflushing the Capillary 00 cece eee eee 54 Performing a Complete System Clean 0 002 55 Printing the Cleaning Log cece eee eee 58 Cleaning Log Options ec cece cece tte ete 59 Clearing the Cleaning Log ce cece eee eee 60 Exporting the Cleaning Log ccc eee ee eee eee eee 60 Viewing and Exporting the Cleaning Event Log 61 Resetting Fluid Levels cece eee eee ees 63 Filling the Cleaning Solution Bottle 0 eee eee 64 Emptying the Waste Bottle 0 65 iv Muse Cell Analyzer User s Guide Cleaning the Capillary 2 02 cece eee eee eee eens 66 Replacing the Flow Cell 0 cee eee ects 67 Preparing the Unit for Depot Service eee ee 69 SeIUDo 2535 ur es ieee ie Aso rob du 71 Setup OPTIONS iusserit n nde e sm CR Y PR eae 71 Users amp Access Levels i wees akg ities Shaina E Eas 73 Adding a New User 0 cee cece eect erent eee 74 Removing a User suasane kreira aeia eee 75 Editing User Information 0 0 cece eee eee ee 76 Recommended Quick Clean 0 eee 76 Printing Layout Options 0 cee eee eee eee eee 77 Editing Favorites eect cho eee Rees REM RP ET RE 78 Managing Instrument Data Storage eee eee eee 80 Exporting by Sere Maui ete t Peete ghee ea had 81 Exporting by Date vi
33. e median values from each triangle System Check 47 Legend Item Description Limit Low A purple line appears 10 below the expected particle count determined by the Check Code Target A green line at the expected particle count determined by the Check Code Limit High A pink line appears 10 above the expected particle count determined by the Check Code 4 Optional You cannot export the trend graph to a CSV file but you can print to a pdf e Select the number of runs last 10 30 or 60 you wish to include from the Trend Graph screen then close the screen Close the System Check Log Options screen e Select the printer tab from the System Check Results screen e Select Print Trend Graph see page 43 then select PDF to USB from the Printer list and finally select Print Viewing the System Check Event Log The system saves a list of all events that occurred during the System Check run to a log file 1 Select Options from the System Check Results screen 2 Select View Event Log The event log contains a list of all events that occurred during the System Check run Every step that the operator performed and every step that the instrument performed independent of the operator are logged as well as warnings and errors that occurred during the run 48 Muse Cell Analyzer User s Guide 3 You can filter the list to view statuses actions errors and or warnings Select the a
34. e spreadsheet file except it has the extension CSV appended to the file name 16 Muse Cell Analyzer User s Guide Event Log Each time you run an assay the system saves a log containing a list of all events that occurred during the assay This information is contained within the FCS data file To view this list of events select Options from the Results screen for the assay then select Event Log mM NOTE There is also an event log for the System Check procedure and the Complete System Clean procedure For information on those event logs refer to the System Check and Cleaning chapters respectively You can filter the list to view statuses actions errors and or warnings Select the appropriate check box es to display the types of events you wish to view A Status A Errors A Action A Warnings 15 Feb 2012 15 38 44 STATUS Acquisi a 15 Feb 2012 15 38 44 STATUS User P 15 Feb 2012 15 38 44 STATUS Assay 15 Feb 2012 15 38 44 ACTION Data Fil 15 Feb 2012 15 38 45 STATUS Waste 15 Feb 2012 15 38 46 STATUS End of 15 Feb 2012 15 38 46 STATUS Establis 15 Feb 2012 15 38 46 STATUS Adjust 15 Feb 2012 15 38 53 STATUS End of 15 Feb 2012 15 38 53 STATUS Waste t 15 Feb 2012 15 38 53 STATUS End of 15 Feb 2012 15 38 54 STATUS Adjust 15 Feb 2012 15 39 12 STATUS Samplir 15 Feb 2012 15 39 12 STATUS Loading 15 Feb 2012 15 39 12 STATUS Purginc 15 Feb 2012 15 39 12 STATUS Purginc WF 15 Feh 2019 15 30 12
35. ecent System Check run This file is used by service personnel to troubleshoot your system if your System Check results continue to fail Use the Export Service Check File feature to export the file so that you can send it to Millipore 1 Select Options from the System Check Results screen 2 Select Export Service Check File 3 Select the USB drive from the Export Location field use My 4GB v GTI 08Dec2011 1 34945 zip ve 4 Enter a name for the log file or leave the default name 5 Select Export 46 Muse Cell Analyzer User s Guide Viewing a Trend Graph You can view a trend graph of the particles mL data from the last 10 30 or 60 System Check runs A data point appears for each of the three replicate values 1 Select Options from the System Check Results screen 2 Select View Trend Graph 3 Choose the number of runs 10 30 or 60 that you wish to plot System Check Trend Graph Particle s ml X 1000 an n 1 Plot Trend for Last 10 Runs ay Last 30 Runs Last60 Runs A legend in the lower left corner lists the information found on the graph A description of each item in the legend appears in the following table Legend Item Description Replicates Outside Range Data point appears as a red triangle value falls outside the high or low 10 limit lines Replicates Within Range Data point appears as a green triangle Replicate Median A black line connects th
36. eck screen or any assay screen 2 Select Reset Fluid Levels from the list of cleaning protocols You can also select Complete System Clean under Essential Tools at the main menu then select View Log Results then Options 3 Select Next The levels are now set to 100 for both the cleaning and waste bottles 1 Empty Waste Bottle and then add 100 bleach to indicator line 7 Ci 2 Fill Cleaning Bottle to indicator line with ICF Load both bottles then select Next to resume assay Cleaning and Maintenance 63 Filling the Cleaning Solution Bottle Fill the cleaning solution bottle with Guava ICF at the start of each day before you run the Complete System Clean procedure and as needed Do not allow the bottle to empty This will pull air into the fluid system and require that you prime the system with water The status bar located at the bottom of the screen indicates the amount of cleaning solution left in the bottle When the remaining volume reaches 20 the value appears in red indicating it is time to fill the bottle with ICF When the volume reaches 1 a message appears indicating that you need to immediately fill the cleaning solution bottle and empty the waste bottle before proceeding User KVT Status Ready for Check DD 23 Example shows cleaning solution bottle has 20 solution remaining 80 has been used 1 Uncrew the tubing from the top of the cleaning solution bottle 2 Press down on
37. elect Export Cleaning Log from the Cleaning Log Options screen 60 Muse Cell Analyzer User s Guide 3 Select the USB drive from the Export Location field 4 Entera file name or use the default file name 5 Select Export 201 2Apr13 1236CleanLog csv 9 A Cancel Export Cancel Export Viewing and Exporting the Cleaning Event Log The system keeps track of and saves a list of all events that occurred during the Complete System Clean procedure to a log file 1 Select Options from the Cleaning Log screen 1 Select Export Event Log from the Cleaning Log Options screen The event log contains a list of all events that occurred during the Complete System Clean Every step that the operator performed and every step that the instrument performed during the procedure are logged Cleaning and Maintenance 61 2 You can filter the list to view statuses actions errors and or warnings 3 26 Jan 2012 14 40 57 26 Jan 2012 14 40 57 26 Jan 2012 14 40 58 26 Jan 2012 14 41 07 26 Jan 2012 14 41 08 26 Jan 2012 14 41 16 26 Jan 2012 14 41 16 26 Jan 2012 14 41 24 26 Jan 2012 14 41 25 26 Jan 2012 14 41 33 26 Jan 2012 14 41 34 26 Jan 2012 14 41 42 26 Jan 2012 14 41 43 26 Jan 2012 14 41 51 26 Jan 2012 14 41 51 26 Jan 2012 14 41 59 26 Jan 2012 14 42 00 STATUS STATUS STATUS STATUS STATUS Waste C STATUS STATUS STATUS Waste Cy STATUS STATUS STATUS End of CI Backflust STATUS
38. ens and materials that come into contact with them can transmit potentially fatal disease To prevent exposure to biohazardous agents follow these guidelines e Handle all biological specimens and materials as if capable of transmitting infection Dispose of waste using proper precautions and in accordance with local regulation Never pipette by mouth Wear suitable protective clothing eyewear and gloves e After emptying waste bottle add bleach to the first fill line It takes approximately 10 mL of bleach to reach the fill line Dispose of waste in accordance with federal state and local regulations Electrical Safety A WARNING Power off the instrument using the Power Options tool on the main menu and disconnect the power cord before replacing fuses Laser Safety The Muse System contains a Class IIIb laser operating at 532 nm in CW mode Light shields within the instrument enclose the path of laser radiation Additionally the instrument enclosure provides secondary protection from any laser radiation Preface 3 A WARNING To avoid exposure to laser radiation or electric shock follow these guidelines e Do not open the instrument or attempt to perform any internal maintenance There are no user serviceable parts e Turn off the power to the system before removing the flow cell Precaution Labels The following labels are affixed to the Muse System di T M TM DANGER u se LASER RADIATION WHEN OPEN Cel
39. f the corresponding bottle Insert the flow cell See Replacing the Flow Cell on page 67 from step step 4 on for details Plug the instrument into a grounded three prong AC outlet When you plug in the instrument for the first time it will automatically turn on Log on as an Administrator See Logging On as the First User on page 29 Muse Cell Analyzer User s Guide 9 Perform a Complete System Clean to prime the fluid system See Performing a Complete System Clean on page 55 10 Perform a System Check procedure See Running a System Check Procedure on page 37 Introduction 7 Muse Cell Analyzer User s Guide System Overview Muse Instrument Overview The Muse Cell Analyzer is a compact instrument for fluorescence based three parameter cell analysis It is designed for easy and intuitive operation and minimal maintenance The integrated touch screen provides an easy to use software interface for operating the system To turn on the instrument press the power button located below the touch screen on the front of the unit To turn off the instrument use the Power Options feature on the main menu displayed on the touch screen Five USB ports one below the touch screen and four on the back of the unit allow you to connect a Muse compatible printer USB flash drive and or keyboard and mouse For a list of approved printers see Approved Printers on page 98 waste bottle cleaning solution
40. f one year from shipment of the products MILLIPORE MAKES NO OTHER WARRANTY EXPRESSED OR IMPLIED THERE IS NO WARRANTY OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE The warranty provided herein and the data specifications and descriptions of Millipore products appearing in Millipore s published catalogues and product literature may not be altered except by express written agreement signed by an officer of Millipore Representations oral or written which are inconsistent with this warranty or such publications are not authorized and if given should not be relied upon In the event of a breach of the foregoing warranty Millipore s sole obligation shall be to repair or replace at its option the applicable product or part thereof provided the customer notifies Millipore promptly of any such breach If after exercising reasonable efforts Millipore is unable to repair or replace the product or part then Millipore shall refund to the Company all monies paid for such applicable Product MILLIPORE SHALL NOT BE LIABLE FOR CONSEQUENTIAL INCIDENTAL SPECIAL OR ANY OTHER DAMAGES RESULTING FROM ECONOMIC LOSS OR PROPERTY DAMAGE SUSTAINED BY ANY COMPANY CUSTOMER FROM THE USE OF ITS PRODUCTS 2012 EMD Millipore Corporation All rights reserved No part of these works may be reproduced in any form without permission in writing Unless otherwise stated in our catalog or other company documentation accompanying the product s our products
41. from the main menu Select Users amp Access Levels from the Setup menu Select the user name then select Edit User Modify the information and select Save Recommended Quick Clean The Recommended Quick Clean feature allows your administrator to select how often a Quick Clean cycle should be performed during a run When a user is running an assay a message appears prompting the user to perform a Quick Clean Only administrators can modify the recommended Quick Cleans 1 Select Setup from the main menu 2 Select Recommended Cleaning Options from the Setup menu 3 Select edit to change the current setting for a given assay The n represents the number of samples that can be run before the user will be prompted to perform a Quick Clean Recommend a Quick Clean every n samples Assay Name n Annexin V amp Dead Cell 20 edit Cell Cycle 12 edit Count amp Viability 20 edit Back User ADM Status 190 100 76 Muse Cell Analyzer User s Guide 4 Use the and buttons to increase or decrease the number of samples between Quick Cleans You can also select to allows users to skip the recommended cleaning for this assay In this case when the cleaning message appears during the assay the user can select Cancel to skip the cleaning 5 Select Apply to apply the changes Recommend a Quick Clean every 20 2X mmt mim Samples Allow user to skip recommended cleaning for this assay Cancel Appl
42. g 80 has been used 12 Muse Cell Analyzer User s Guide Main Menu The main menu allows you to choose an assay or an Essential Tool Navigation bar Muse Setup Title bar with Setup and tube eject buttons Favorites ta Count amp Viability Annexin V amp Dead Cell Select an assay Cell Cycle Essential Tools System Complete Power 08 MAR 2012 12 29 User ADM Status wes fyror Below the navigation bar is the title bar containing the Setup button and eject button The eject button releases the sample loader arm allowing you to manually unload a tube Setup which is accessible from the main menu only allows you to set specific system features such as managing users and data and setting specific software options See Setup on page 71 for details on the Setup menu j Muse sep v Title bar The lower part of the screen is divided into two main sections Favorites assays and Essential Tools Favorites allows you to select an assay to run Essential Tools allows you to check the system performance clean the system and power off or log off of the system System Overview 13 Favorite Assay Buttons The main menu allows you to select from the following assays e Count amp Viability for performing cell counting and viability assays e Annexin V amp Dead Cell for performing apoptosis assays e Cell Cycle for performing cell cycle assays F
43. h Cell Concentration warning during acquisition If the concentration of the stained cell sample for acquisition is high 5500 cells uL the accuracy of the data will most likely be compromised Repeat sample preparation with a higher dilution factor to allow for adequate cell numbers 88 Muse Cell Analyzer User s Guide Problem Recommended Solution Background staining and or non specific staining of cells The cells may be damaged as dead cells tend to aggregate and non specifically bind fluorescent reagent Avoid damaging the cells when handling and processing them in culture Low level of staining Although the assay procedure was optimized for multiple cell types every cell line behaves differently A low signal may indicate that the cell concentration may be too high for the amount of reagent use Restain cells at a lower concentration Ensure proper controls are used Variability in day to day experiments e f the results are inconsistent check that the samples were well mixed prior to acquisition Cells may quickly settle in your samples and your results will be inaccurate unless the cells are mixed just prior to acquisition Monitor experimental cell cultures to ensure that cell viability and cell numbers being analyzed are consistent Any drop in cell numbers or viability can influence experimental results e f there appears to be day to day variation of the staining pa
44. he capillary by simply loading and unloading a tube of water Use this option to avoid carryover 1 2 3 4 5 Select Clean from the System Check screen or any assay screen Select Capillary Rinse from the list of cleaning protocols Load a full tube of DI water on the unit Select Eject Load sample tube with DI water to rinse the capillary then tap Eject Raise Load DI Water Sample Loader Select Eject LC y Us Cancel ENS UM Cancel amp Eject Rinse Capillary Discard the tube of water Running Quick Clean Quick Clean is a short cleaning cycle that cleans the fluid system during and after an assay or as often as you like throughout the day If you are running lots of samples with high background run Quick Clean according to the frequency recommended for the assay a Recommended Quick Clean message will appear for every N number of samples 1 2 3 Select Clean from the System Check screen or any assay screen Select Quick Clean from the list of cleaning protocols The system prompts you for ICF however you can load a tube containing any one of the following solutions e DI water to quickly flush out the system e Guava ICF to clean the system follow with a second Quick Clean using water to rinse Cleaning and Maintenance 53 e 10 bleach solution in Guava ICF 1 part bleach in 9 parts Guava ICF follow with a second Quick Clean using water to rinse 4 Select Clean to run t
45. he solution Load sample tube containing DI Water Load DI Raise Water Sample Loader Select Clean 5 Ifyou used water you are done with the Quick Clean procedure If you used Guava ICF either straight or with bleach you will need to run water to rinse the fluid system Select Quick Clean from the cleaning protocol screen Load a full tube of DI water then select Clean You may continue running samples or leave the tube of water loaded on the unit until you are ready to use the system again Backflushing the Capillary The Backflush feature reverses the flow of fluid out of the capillary Use this feature when you suspect that the fluid pathway is clogged 1 If you suspect a clog during acquisition select Clean from the System Check screen or any assay screen 2 Select Backflush from the list of cleaning protocols 3 Load a tube containing 100 uL of 20 bleach 54 Muse Cell Analyzer User s Guide 4 Select Clean Load sample tube with 2096 bleach to backflush the system Load Raise Select Clean 20 bleach Sample Loader Select Clean Xr Cancel Clean Cancel amp Eject Start Backflush 5 When the backflush is complete throw out the tube of bleach you used in step 3 as it may contain debris from the backflush 6 Select Quick Clean to rinse the residual bleach from the capillary Load a full tube of DI water on the unit and select Clean 7 Replace the sample and continue with the assay or p
46. heck Results screen 2 Select View Event Log 3 Select Export from the Data Set Event Log screen System Check 49 4 Select the USB drive from the Export Location field USB1 My 4GB 2011Dec08_141423 log 5 Enter a name for the log file or leave the default name 6 Select Export 50 Muse Cell Analyzer User s Guide Cleaning and Maintenance The system s sampling precision depends on the integrity of the fluid pathway therefore it is important to maintain a clean system Do not allow samples to remain loaded on the instrument for extended periods of time as they may eventually clog the fluid system Perform frequent cleaning cycles to prevent the build up of cellular debris that may restrict sample flow Cleaning Protocols Several cleaning options are available depending on the type of cleaning you wish to do The following protocols are available by selecting Clean from any assay screen as well as the System Check screen shown below Run Results Select Clean from System Check or any assay screen to access the cleaning options e Capillary Rinse allows you to rinse the outer surface of the capillary This can be helpful if you want to reduce carry over between samples or when cell counts are critical e Quick Clean is a short cleaning cycle that allows you to clean the system during and after an assay or as often as you like throughout the day e Backflush reverses fluid out
47. iability e Annexin V amp Dead Cell e Cell Cycle Refer to the assay user s guide for detailed instructions on preparing samples and running an assay using the assay specific software module Muse Cell Analyzer Introduction 5 Setting Up Your System Your Muse System is easy to install Simply plug it in and start operating Components Your Muse System shipped with the following components Muse Cell Analyzer and power cord cleaning and waste bottles and corresponding color coded tubing flow cell USB flash drive with software recovery files and user s guide Unpacking the Instrument Follow the instructions in the Quick Start Guide to unpack and set up your instrument Here are the steps outlined briefly 1 Remove the instrument from the shipping box You may wish to save the box and foam insert in the event the instrument needs to be moved to a different location Place the unit on a stable surface Although it is compact and portable it contains precisely aligned optical components that are sensitive to jarring movements Connect the color coded fluid tubing to the back of the instrument Add 10 mL of bleach to the waste bottle and fill the cleaning solution bottle to the indicator line with Guava ICF Place the fluid bottles in their respective receptacles Match the icon on the fluid bottle with the icon in the receptacle Connect the fluid tubing from the back of the instrument to the top o
48. ings from your own data folders in the Muse directory and USB drive Assay Instrument Settings File Name Count amp Viability filename VIA GST Annexin V amp Dead Cell filename NEX GST Cell Cycle filename CCY GST You can recall this file later to e download the instrument settings to the instrument for acquisition e apply the gates and markers to data during acquisition System Overview 21 Retrieving Instrument Settings 1 From the Settings screen at the start of a run select Retrieve Settings Se d Retieve setings Mix and Load Sample simil Teper Select Run Ui Oy Oe Cancel Run Cancel amp Eject etting 2 Select the location where the settings file is stored 3 Select the file and then select Retrieve You can filter the list by date Select the calendar and choose the date when the settings were saved Or if you know the name of the file enter any part of it in the Filter text field ln ccu E cie i o orocasc u4d Retrieve Settings Retrieval Location MUSE 4C Praqram Files Millipare Marse M Filter PT 15FEB2012 163431 Cancel Cancel Retrieve 22 Muse Cell Analyzer User s Guide The settings are downloaded to the instrument The system prompts you to run the adjust settings step e Select NO if you wish to skip the adjust settings step and use the settings you just retrieved to run the assay This will advance you to the Sample Info
49. lect Import Select USB thumb drive E Muse Q us My 4GB g Include Public Folder E Public User ADM Status Ge 100 l 6 The system automatically searches the drive for the user data A message appears indicating that any data currently on the system will Setup 83 7 be overwritten Select YES to proceed or Cancel to cancel the importing Home gt Setup gt Manage Instrument Storage Import All User Data This will overwrite this instrument s data with information from USB1 My 4GB Proceed Select Close to the message that data was successfully imported Exporting All User Data Use the Export All User Data feature to back up all data files Only administrators can export all user data 1 Select Setup from the main menu 2 Select Manage Instrument Storage from the Setup menu 3 Select Export All User Data 4 Optional If you wish to delete the data from the system after it s exported select Delete files from the instrument after successful export 5 The system displays the drive space needed for the export and displays only the USB drives with the available space Select the USB drive and then select Export 84 Muse Cell Analyzer User s Guide BI NOTE This could take several minutes depending on how much data you are exporting Select USB thumb drive USB1 My 4GB Delete files from instrument after successful export EN User KG Status we fes
50. low channel on the Muse Cell Analyzer can measure fluorescent light from dyes that emit signals similar to PE Glossary 93 94 Muse Cell Analyzer User s Guide Ordering Information For ordering information contact the nearest Millipore office by calling 1 800 645 5476 or visiting us on our website at www millipore com offices Millipore and its distribution network will provide Muse products to all sectors of life science research in certain countries outside North America and Europe Part Catalog Number Muse Cell Analyzer 0500 3115 cleaning solution bottle and corresponding fluid tubing 0110 7865 waste bottle and corresponding fluid tubing 0110 7860 flow cell assembly 0500 3120 flow cell cleaning tool 6000 2820 1 5 mL tubes Eppendorf quantity 500 1000 0785 Muse Resource flash drive software and user s guide 0110 7880 Muse Cell Analyzer User s Guide 0110 7895 Reagents for Muse System Catalog Number Muse System Check Kit 100 tests MCH100101 Muse Count amp Viability Kit 100 tests MCH100102 Muse Count amp Viability Kit 600 tests MCH600103 Muse Count amp Viability Kit 200X MCH100104 Muse Annexin V amp Dead Cell Kit 100 tests MCH100105 Muse Cell Cycle Kit 100 tests MCH100106 Muse Cell Dispersal Reagent 100 tests MCH100107 Ordering Information 95 96 Muse Cell Analyzer User s Guide Muse System Physical Characteristics instrument weight instru
51. luid ICF for system cleaning For information on filling the bottle refer to Filling the Cleaning Solution Bottle on page 64 The waste bottle captures the sample fluid after it exits the fluid system Add 10 mL of bleach to the waste bottle after you empty it and before installing it in the unit For information on emptying the waste bottle refer to Emptying the Waste Bottle on page 65 Fluid System Sample uptake occurs through the capillary part of the flow cell assembly Sampling is regulated by a variable speed fluid pump The pump does not require sheath fluid or supplementary fluids for sample acquisition capillary Because the system s sampling precision depends on the integrity of the fluid pathway it is important to maintain a clean system Do not allow samples to remain in the capillary for extended periods of time as they may eventually clog the system Perform frequent cleaning cycles to prevent the build up of cellular debris that may restrict sample flow If a clog does occur you can clear it by using the backflush feature which reverses the flow of fluid and flushes it out of the capillary at high speed See Backflushing the Capillary on page 54 for detailed instructions on using the backflush feature At the start of each day perform a Complete System Clean to prime the fluid system See Performing a Complete System Clean on page 55 Always leave a tube of deionized water on the instrument when not in
52. me User ADM Status Ge fees 30 Muse Cell Analyzer User s Guide 3 Use the arrow buttons above and below each field to select the day month year hours and minutes Set Date and Time Date Monday 06 FEB 2012 4 Select Apply 5 See Adding a New User on page 74 for information on adding new users Getting Started 31 Logging On Once Users are Added When you start the system a screen appears allowing you to select your user name from a list 1 Select your name from the list to display the main menu If your name does not appear contact your system administrator Katie Gillis Patricia Toth Julie Clor Guest n User Status d 100 Syros 32 Muse Cell Analyzer User s Guide 2 Enter your PIN and press OK If security is not an issue and you choose not to enter a PIN each time you log in you can select Remember User PIN The next time you log in simply select OK to accept the PIN Username Katie Gillis i Remember User PIN The main menu appears after your PIN is correctly entered Forgetting Your User PIN If you forget your PIN your administrator can reset it by assigning you a new PIN If you are an administrator and you forget your PIN another administrator can reset it If you are the sole administrator and you forget your pin you will no longer have administrator access to the system and therefore cannot perform administrator functions
53. ment 5 Release the clamp securing the flow cell and carefully remove the flow cell 6 Use an empty syringe to flush the flow cell with air Place the flow cell in its original box 7 Lift the loader arm in the up position 8 Place the shipping insert with the foam attached on a flat surface then place the instrument in the foam cut out The tall block of foam should be on the right as it will be positioned under the loader arm when sides are folder up 9 Fold the sides of the insert up making sure the foam block fits under the loader arm 10 Place the power supply and power cord behind the instrument 11 Use both handles on the insert to lift the instrument and lower it into the shipping box Cleaning and Maintenance 69 12 Place the fluidics box with flow cell fluid bottles and tubing in the space in front of the instrument 13 Ship the instrument to EMD Millipore ATTN Muse RMA insert the RMA number issued to you 25801 Industrial Blvd Hayward CA 94545 70 Muse Cell Analyzer User s Guide Setup Options Setup contains the tools necessary to set various system features such as adding new users and setting their access levels selecting how often users should clean during an assay and choosing printer and file storage options Select Setup from the main menu Setup 2 Select the option you wish to perform M NOTE Depending on your user access level some options may not be available to you
54. ment size height width depth connectors Operating Environment temperature external power supply input voltage range main unit input voltage fuse rating Optics laser forward scatter detector fluorescence detectors Signal Processing parameter dynamic range pulse processing time Specifications 13 1 Ib 5 94 kg 8 69 in 22 07 cm 8 12 in 20 62 cm 11 11 in 28 22 cm power Kycon KPJX PM back panel for motherboard Intel Db25MW five USB connectors one on front and four on back of instrument 16 35 C 60 95 F 100 240 VAC 50 60 Hz 80 W 15VDC 5A auto resettable 532 nm green laser photodiode photodiodes YLW 576 28 RED 680 30 4 0 decade digital signal processing every particle time stamped Specifications 97 Fluidics flow cell dimension pump samp cleani e flow rate waste generation dead samp samp volume e concentration e requirement Data Management comp uter data file structure Perfor mance counting accuracy counting precision Approved Printers ng waste bottles rectangular capillary 1 5 mm x 0 8 mm with 100 pm round bore positive displacement 7 L min to 36 uL min 50 mL plastic co polyester bottles with screw tops lt 40 mL 50 uL final particle concentration of 1 x 10 to 5 x 10 particles mL as few as 2 000 cells test typically 25 000 100 000 cells test depending on the assay
55. mp Viability e Annexin V amp Dead Cell e Cell Cycle Home Assay Settings Essential Tools System Complete Power Check System Clean Options 08 MAR 2012 12 29 User ADM Status wes M100 For information on performing a System Check see Running a System Check Procedure on page 37 For information on running an assay refer to the kit user s guide which can be found at www millipore com muse 36 Muse Cell Analyzer User s Guide System Check The System Check procedure is used to verify the performance of your Muse System by assessing counting accuracy and fluorescence detection The Muse System Check Kit contains Muse System Check Beads and Muse System Check Diluent Perform the System Check procedure daily before running any of the assays to ensure that the instrument provides reliable accurate results Running a System Check Procedure Run a System Check at the start of each day that you use the instrument to ensure that it is performing properly Three replicates of the System Check Bead sample are acquired The results are averaged to determine if they are within the expected range Run a Complete System Clean at the start of each day and before performing the System Check procedure A cleaning cycle will prime the fluid system and remove bubbles that may have formed in the tubing See Performing a Complete System Clean on page 55 for details 1 Prepare a 1 20 dilution of S
56. ng and discrimination of viable and nonviable cells for a variety of difficult cell sample Muse Annexin V amp Dead Cell Kit Used to assess early and late apoptosis The assay relies on the translocation of phosphatidyl serine PS to the outer surface of the cell membrane an event often associated with the onset of apoptosis Muse Cell Cycle Kit Identifies and measures the number of cells within the various phases G0 G1 S and G2 M of the cell cycle Muse Cell Dispersal Reagent Enzymatic reagent that gently disaggregates clumped cells in suspension improving the accuracy and precision of cell counts The formulation of CDR has been optimized for use with Muse Count amp Viability Reagent Preface 1 About this Guide The Muse Cell Analyzer User s Guide provides detailed information on operating and maintaining the Muse System This guide provides instructions for setting up the system managing users and data files running a System Check procedure and cleaning the system It does not include instructions for using the software modules to acquire samples and analyze data For information on preparing samples and running them on the instrument refer to the specific kit user s guide located at www millipore com muse Conventions Used in This Guide BI NOTE Points out additional information that may be helpful A WARNING Alerts you to situations that could result in bodily harm instrument damage failure in a pr
57. ocedure or incorrect results Help 1 Read through the section of the guide specific to the operation you are performing Refer to the table of contents and index to locate information A glossary is included to assist you with any unfamiliar terms 2 See the troubleshooting section for a list of problems and suggested solutions 3 Refer to the technical support contact information below e For ordering information or technical support call toll free in the USA and Canada Phone 1 800 MILLIPORE 645 5476 Fax 1 800 645 4539 e Outside the US visit www millipore com offices for up to date worldwide contact information 4 For information on performing an assay refer to the specific kit user s guide which can be found at www millipore com muse 2 Muse Cell Analyzer User s Guide Safety The Muse System is equipped with safety features for your protection Use the system only as directed in this user s guide Do not perform instrument maintenance or service except as specifically stated Read the following safety information before using the system General Safety A WARNING If this instrument is not used in the manner indicated by the instructions in this guide the safety features of the instrument may be impaired Follow these guidelines e Use only the tubes specified The use of tubes other than those specified may result in damage to the instrument Biological Safety A WARNING All biological specim
58. of the capillary Use this feature if you suspect a clog Always load a tube of 20 bleach when you perform a backflush e Complete System Clean allows you to thoroughly clean the instrument at the end of the day or between assays if needed Cleaning and Maintenance 51 e Reset Fluid Levels sets the fluid level status for the waste bottle and cleaning solution bottle back to 100 and 100 after you have emptied the waste bottle and filled the cleaning solution bottle Select a cleaning option Capillary Rinse Rinse outer surface of capillary Quick Clean Rinse fluid path Cleaning options Backflush Clear fluid path Complete System Clean Reset Fluid Levels Reset Waste and Cleaning Bottle Levels Close Close Window Cleaning the Outside of the Unit Clean the outside of the instrument by wiping it down with a soft cloth moistened with a 70 isopropyl alcohol or a 10 bleach solution in DI water Follow with a cloth moistened with water To avoid getting excess liquid on the instrument do not spray these solutions on the instrument To clean the touch screen wipe it with 70 isopropyl alcohol A WARNING To avoid exposure to laser radiation or electric shock do not open the unit or attempt to perform any internal maintenance There are no user serviceable parts 52 Muse Cell Analyzer User s Guide Rinsing the Capillary Capillary Rinse allows you to rinse the outside of t
59. ons starting on page 45 44 Muse Cell Analyzer User s Guide System Check Options Select Options at the top of the System Check Results screen to export or clear the System Check results log export the Service Check file or view the trend graph or event log Syst em Check m Optio ns Clear Check Log Export Check Log Export Service Check File View Trend Graph View Event Log Close Close Window Clearing the System Check Log The System Check Log is a list of all past System Check results After exporting the System Check Log you may wish to delete all entries in the log Once the data is cleared you will no longer be able to view past results or generate trend graphs Only administrator level users can clear the System Check Log 1 Select Options from the System Check Results screen 2 Select Clear Check Log 3 Select Clear to confirm System Check 45 Exporting the Check Log The System Check Log is a list of all past System Check results Only administrator level users can export the System Check Log 1 Select Options from the System Check Results screen 2 Select Export Check Log 3 Select the USB drive from the Export Location field USB1 My 4GB v 2011 Dec08_133422 csv 4 Enter a name for the log file or leave the default name Select Export Exporting the Service Check File The Service Check file is a zipped file containing the detailed results from the most r
60. played as well as the amount of storage being used For more information on managing instrument storage see Managing Instrument Data Storage on page 80 Home gt Sotup gt Manage Instrument Storage Maximum available storage 160GB Current storage usage 45GB Available and used space Appending Files You can open an existing data set and append data to this file 1 At the start of the assay select View Results from the assay screen The Retrieve Data Set screen appears Retrieve Data Set Retrieval Location MUSE C IP ram Files Millipari Musie MUSE Pat PT 23MAR2012 152619 PT 23MAR2012 152302 20 Muse Cell Analyzer User s Guide 2 Select the location where the file is located Keep in mind that you will not have access to the files from other users unless the files were exported to the Public folder 3 Use the Filter field to sort by the file name or any part of the name Or use the calendar to search by date Touch the calendar to open it and select the date 4 Select the file and then select Retrieve Instrument Settings The system allows you to save instrument settings files These files contain instrument settings as well as gates and marker settings The extension GST is appended to the file name You can retrieve these files from the MUSE directory the Public folder or USB drive when you start an assay If you are an operator level user you can only access instrument sett
61. ppropriate check box es to display the types of events you wish to view m Warning A Action A Error 07 Dec 2011 16 12 23 STATUS 07 Dec 2011 16 12 23 STATUS 07 Dec 2011 16 12 23 STATUS 07 Dec 2011 16 23 23 STATUS 07 Dec 2011 16 23 23 STATUS 07 Dec 2011 16 23 23 STATUS 07 Dec 2011 16 23 23 STATUS 07 Dec 2011 16 23 23 STATUS 07 Dec 2011 16 23 25 STATUS 07 Dec 2011 16 23 26 STATUS Analysis j User 4Z AssayNC Acquisiti User 4Z Assay Ni 07 Dec 2011 16 23 23 ACTION 07 Dec 2011 16 23 23 STATUS Data File Sampling AdjustSe Waste C 07 Dec 2011 16 23 23 STATUS 07 Dec 2011 16 23 23 STATUS 07 Dec 2011 16 23 23 STATUS 07 Dec 2011 16 23 25 STATUS Waste C 07 Dec 2011 16 23 26 STATUS AdjustSe 07 Dec 2011 16 23 26 STATUS End of V Establist Setting T Threshol End of V Ready fc gt Close Close Window Filter the event log list by selecting the types of events you wish to view If errors or warnings occur during a run a message appears in red on the results screen indicating that errors warnings have been logged For a list of System Check errors and warnings see page 17 Exporting the Event Log You can export the System Check event log to a CSV file If you are currently viewing the event log proceed to step 3 The entire event log is exported regardless of whether you filtered the list 1 Select Options from the System C
62. ries in the log list While only administrator level users can clear the log any user can export the log Select Options from the Cleaning Log screen If you did not just complete the system cleaning procedure select Complete System Clean under Essential Tools at the main menu then Cleaning and Maintenance 59 select View Log Results from the Complete System Clean screen then Options Clear Cleaning Log Export Cleaning Log Export Event Log Reset Fluid Levels Reset Waste and Cleaning Bottle Counts i Cancel 2 Referto the following sections for information on the task you wish to perform Clearing the Cleaning Log After you export the Cleaning Log you can clear the log Only administrator level users can clear the Cleaning Log 1 Select Options from the Cleaning Log screen 2 Select Clear Cleaning Log from the Cleaning Log Options screen A message appears informing you that the action cannot be undone 3 Select Clear to confirm and clear the log Exporting the Cleaning Log You can export the cleaning log to a CSV file The file contains a list of all Complete System Clean procedures performed since the last time the cleaning log was cleared The list includes the dates and times that each Complete System Clean was run as well as the operators who performed the cleaning procedure and whether the cleaning was completed or aborted 1 Select Options from the Cleaning Log screen 2 S
63. rocedure you were running Performing a Complete System Clean Run the Complete System Clean feature to clean the unit at the end of each day or between assays if a thorough cleaning is needed You can also run this cleaning cycle to prime the fluid system or if you suspect there is air in the fluid lines Always ensure the cleaning solution bottle is filled with ICF whenever you run the Complete System Clean 1 Select Complete System Clean under Essential Tools at the main menu This feature is also available by selecting Clean from the System Check Screen or any assay screen then selecting Complete System Clean from the list of cleaning protocols Cleaning and Maintenance 55 2 Select Run Complete Clean Complete System Clean ow Li Run Complete Clean View Results Last Complete Clean 19 JAN 2012 03 07 PM Displays the last time a complete system clean was performed luser ACM Status Rewir tor Charing B10 yes 3 Load a full tube of Guava ICF on the unit and select Run Run ICF Step 1 of 2 Load sample tube with ICF then select Run to begin the cleaning Raise 56 Muse Cell Analyzer User s Guide 4 When the ICF cycle is complete the system prompts you for a tube of DI water Load a full tube of DI water on the unit and select Continue Run DI Water Step 2 of 2 Load sample tube with DI water then select Continue to begin the cleaning Raise Load D
64. splays a PASS FAIL result and the Particles mL value for each replicate as well as the average Options View past System Check results Bead Info Bead lot date and time PASS FAIL result Sample Particles Bead Lot 110279 Expiration Date 06 Jan 2012 Check Code 6543290 Finish Next Run Return to Launcher Status field The bead lot number date and time of the run appears just above the Bead Info and printer tabs All runs in the System Check log are displayed in this format The Status field at the bottom of the screen indicates that the System Check was completed System Check 41 System Check Results The result appears as PASS or FAIL The bead count Particles mL for each replicate and the average are displayed as well as the CV If any result for Particles mL falls outside 10 of the expected value the result is outside the acceptable range and appears in red If the procedure fails touch the Help button to display troubleshooting information BI NOTE The System Check procedure can also fail if the red and yellow mean fluorescence intensities MFls fall outside the expected results If all Particles mL values are green but the result is FAIL contact Millipore Technical Support Touch Help button to get troubleshooting information Bead Lot 12 0070 Expiration Date 01 Mar 2015 Check Code AZRSR99D Finish Next Run Return to Launcher F i K User
65. ss down on the waste bottle to release it from the unit The bottle will pop up slightly allowing you to remove it Push down on the bottle to remove it Carefully unscrew the cap 4 Empty the contents according to your local and state biohazardous waste disposal guidelines 5 Rinse the bottle with water Add approximately 10 mL of bleach to the empty waste bottle Cleaning and Maintenance 65 7 Replace the cap and reinstall the bottle in the instrument Align the trash can icon on the bottle with the notch line on the instrument Press down on the bottle to engage it Reconnect the fluid line to the top of the bottle Cleaning the Capillary Use the flow cell cleaning tool syringe to clean the flow cell 1 Use the eject button on the main menu or any assay screen to lower the sample loader arm 2 Open the flow cell access door at the top of the instrument 3 Use your fingers to unscrew the tubing from the instrument 4 Fill the syringe with DI water or Guava ICF then screw the syringe onto the tubing 5 Apply gentle steady pressure to the plunger and watch as the fluid flows from the tip of the capillary e Make sure the fluid stream is straight If it is not straight the tip of the capillary may be chipped or there may be a partial clog in the flow cell 66 Muse Cell Analyzer User s Guide oO c e Check the capillary to ensure there are no leaks along the length of the tube or liquid pooling a
66. t Bead Info ae Enter the following information Bead Lot 78965 Exp Date stMa2oi2 S Touch a field to display a keypad allowing you to enter the values Check Code AWRRR999 Back Cancel Check User KG Status Ready tor System Check Dor fe 7 Mix the tube of prepared beads and load it on the system 8 Select Run Run 1st Replicate ow Mix and Load Raise Sample Sample Loader Select Run mq Gance un 5 Fiery The system performs a prime then acquires the first replicate The progress bar and fly wheel provide indicators as to the status of acquisition The progress bar is divided into three sections one for each replicate If the fly wheel is turning but the progress bar in not System Check 39 advancing the fluid system may be clogged or the beads may have settled to the bottom of the tube If the beads settled select Abort unload the tube and mix Then reload and select Run System Check again System Checking Sample Particles Info iml 965 Progress bar gives an estimate of the acquisition status for each replicate Fly wheel lets you know the instrument is acquiring sample 9 Remove the tube and vortex it to resuspend the beads 10 Load the tube and select Run The system acquires the second replicate 40 Muse Cell Analyzer User s Guide 11 Repeat steps 9 and 10 to acquire the third replicate Upon completion the system di
67. t the top of the flow cell s Unscrew the syringe tool from the tubing Reconnect the tubing to the instrument Make sure the tubing is screwed on tightly and there are no constricting kinks or twists Close the access door Prime the system by running a Quick Clean procedure Perform a System Check procedure Replacing the Flow Cell You can replace the flow cell if it becomes damaged or clogged so severely that backflushing and cleaning the system do not fix the problem When removing and replacing the flow cell handle it with care The capillary tube is fragile avoid touching it unnecessarily Do not force the flow cell into the receptacle A WARNING To avoid exposure to laser radiation turn off the power to the Muse System before attempting to remove the flow cell Use the eject button on the main menu or any assay menu to lower the sample loader arm Open the flow cell access door at the top of the instrument Cleaning and Maintenance 67 3 Use your fingers to unscrew the tubing from the instrument 4 Release the clamp that holds the flow cell assembly in place Press down on the left edge of the clamp to release the forked side on the right 5 Holding onto the tubing as closely to the green connector as possible gently pull up on the flow cell assembly to remove it from the receptacle Keep the assembly completely vertical until the capillary is clear of the instrument 6 Discard the flow
68. ted 27 3 Fill the cleaning solution bottle to the fill line with Guava ICF Empty the waste solution bottle then add bleach to the first fill line approximately 10 mL See Filling the Cleaning Solution Bottle on page 64 and Emptying the Waste Bottle on page 65 for details Whenever you fill the cleaning solution bottle and empty the waste bottle you will need to reset the fluid levels Resetting the fluid levels sets the fluid indicators in the status bar to 100 and 100 See Resetting Fluid Levels on page 63 for information E User ADM Status Cleaning Complete m 100 im 100 Run a Complete System Clean at the start of each day to prime the fluid system See Performing a Complete System Clean on page 55 Turning Off the Unit 1 Run the Complete System Clean procedure at the end of the day before shutting down the unit See Performing a Complete System Clean on page 55 for details Leave the tube of DI water on the sample loader WARNING Do not leave a tube of Guava ICF bleach or any other cleaning agent loaded on the instrument overnight or for an extended period of time Prolonged exposure to strong oxidizing agents will damage the flow cell Always leave a fresh tube of DI water on the system when shutting it down Change the tube of water regularly to ensure it is clean and free of particles Select Power Options from the main menu under Essential Tools Essential Tools System Complete Power Che
69. tem Administrators have privileges to all options within the Setup menu Operators have the following privileges only e ejecta USB drive e adjust the volume e edit their own user name and or PIN e select the default printer e edit favorites assay list that appears on the main menu e export their own data by user or date however cannot delete after exporting e export a Service Check file e export the cleaning log Tasks that operator level users cannot perform are e change the cleaning options e import or export all user data e edit printing layout options e update the software e export the System Check log e export data or save instrument settings to the Public folder Setup 73 To Access the Users amp Access Levels Screen 1 Select Setup from the main menu 2 Select Users amp Access Levels from the Setup menu The Users amp Access Levels screen appears Administrators can add and remove users Any user can edit their name and PIN Jsers amp Access Levels Initials User Name Access Rights GST Guest Guest Add New Remove Edit User pa User User Back User ALM status Bs fons 3 Refer to the following sections for more information on adding removing and editing user information Adding a New User Only administrators can add new users to the system When you add the first new Administrator user the default Administrator ADM will be deleted 1 Select Setup from the
70. the cleaning solution bottle to release it from the unit The bottle will pop up slightly allowing you to remove it Push down on the bottle to remove it 3 Unscrew the cap 4 Fill the bottle to the fill line with Guava ICF 5 Replace the cap and reinstall the bottle in the instrument Align the shower icon on the bottle with the notch line on the instrument Press down on the bottle to engage it 6 Reconnect the fluid line to the top of the bottle 64 Muse Cell Analyzer User s Guide Emptying the Waste Bottle Empty the waste bottle at the start of each day before you run the Complete System Clean procedure and as needed The status bar located at the bottom of the screen indicates the amount of space left in the waste bottle When the remaining space reaches 20 the value appears in red indicating it is time to empty the waste bottle When the remaining space reaches 1 a message appears indicating that you need to immediately empty the waste bottle and fill the cleaning solution bottle before proceeding User KVT Status Ready for Check ni 20 96 my 22 Example shows waste bottle has 20 capacity remaining 80 full A WARNING Handle all biological specimens and materials they come in contact with as if capable of transmitting infection Dispose of these materials using proper precautions in accordance with federal state and local regulations 1 Unscrew the tubing from the top of the waste bottle 2 Pre
71. ttern ensure the Muse Cell Analyzer is working properly Run the Muse System Check procedure to verify proper instrument function and accuracy e Always monitor threshold settings especially if using different cell types to ensure cell events are not excluded Troubleshooting 89 90 Muse Cell Analyzer User s Guide acquisition analysis coefficient of variation CV data set detector dot plot FCS file Glossary The electronic and software function of collecting various types of information from a cell sample The software function of numerically and graphically manipulating data to identify and separate cell populations for the purpose of calculating relevant statistical information The ratio of the standard deviation to the mean expressed as a percent It is calculated using the formula CV 22 x 100 X A series of samples included within one file for a selected assay An FCS file and a spreadsheet file are saved for each data set A device used to measure light intensity The fluorescence detectors are photodiodes yellow and red The FSC detector that measures cell size is also a photodiode Both output a current that is proportional to the intensity of incident light A graphical representation of two parameter data Each axis of the plot displays values for one parameter A dot represents the values for a cell or particle Flow Cytometry Standard file A data file containing
72. vice For assay specific troubleshooting refer to the assay specific kit user s guide at www millipore com muse Problems Problem Recommended Solution During startup computer freezes on particular screens Cycle power by pressing the power button located below the touch screen Message Instrument could not be detected Cycle power If message appears after rebooting contact Technical Support Instrument settings could not be retrieved Ensure settings are located in the correct user folder If settings are to shared among user ensure settings files are saved to the Public folder Forgot user PIN See Forgetting Your User PIN on page 33 or contact Tech Support Continued System Check failures after Complete System Clean Ensure the cleaning solution bottle is full of ICF and all connections are finger tight Loader arm is stuck in the up position Eject button on screen does not lower it Use the loader arm release lever in the flow cell hatch See Loading and Unloading a Tube on page 10 Loader arm is not dropping loading Ensure the loader arm release lever in the flow cell access door is in the proper position see page 10 Troubleshooting 87 Problem Recommended Solution System Check failures one ore more Particles mL results fall outside the acceptance range appears in red 1 Ensure the correct Bead Lot and Check Code
73. who performed the cleaning procedure and whether the cleaning was completed or aborted 1 Select the printer tab from the Cleaning Log screen If you did not just complete the system cleaning procedure select Complete System Clean under Essential Tools at the main menu then select View Log Results from the Complete System Clean screen Optional Select Print Preview to see the list as it will print If necessary select the printer from the Printer drop down list If a printer is detected it will appear as the default If no printers are available the default is PDF to USB allowing you to save the results to a PDF file Select Print to print the list If no printers are available or if you wish to export to a PDF select PDF to USB then select Print The Export to PDF screen appears Select the 58 Muse Cell Analyzer User s Guide USB port enter a file name for the report file or leave the default name Select Export 10 OCT 2011 10 29 14 Printer EEG HP 675 All in One v FS TS Status Ready Paper Size 8 x 11 Print Cleaning Log Print Preview ELM 2 l 7 Finish Next Cleaning Return to Launcher Run Next Cleaning user er Status Cleaning Complete Results FER EEE Cleaning Log Options Printer tab Select to print the log Or select Export to save to a PDF You can view the cleaning log and clear or export the ent
74. x de pria a e eE EE RE IRE 82 Importing All User Data sssssssss SR 83 Exporting All User Data 00 eee eee eee eee 84 Software Update cece 85 Service Tools amp ccr ce tere fee tse b Yes 86 Troubleshooting nici serie Rr RRRPIPREEAREEE ERI 87 Problems 5 uleteu esap E ice et mtra ot unte eur enel oe ac s 87 Glossary 02s eos iti nai su senteee bees Se Genee aaa 91 Ordering Information leen 95 Sp ciTicatiOHS duret hectic n ERE a unb i i E ENS 97 vi Muse Cell Analyzer User s Guide Preface The Muse System is the first compact easy to use desktop cell analysis system that can perform a wide range of cellular assays Assay and Reagent Overview For information on performing the assay protocols refer to the specific kit user s guide which can be found at www Millipore com muse For order information see page 95 For Research Use Only not for use in diagnostic procedures Muse System Check Kit Verifies the performance of the system by assessing counting accuracy and fluorescence detection using a standardized fluorescent bead reagent The kit contains a bead reagent and diluent Muse Count amp Viability Kit Used to determine viability and total cell count Accurate assessments can be made with a wide variety of cell lines even those with unusual culture conditions or a tendency to aggregate Muse Count amp Viability Reagent 200X Enables accurate counti
75. y Discard Changes pe ADM Status Bie fy Printing Layout Options Any user can select the default printer Only administrators can edit the layout options 1 Select Setup from the main menu 2 Select Printing Layout Options from the Setup menu 3 Select the printer you wish to use as the default 4 Select the layout options you wish to use e Include Signature Field adds a signature field to the printout e Include Plots includes plots on the printouts Setup 77 BI NOTE Only administrators can edit the layout options Printer Microsoft Office Document Image Writer Connected Ready e Connected Ready ix Layout Options g Include Signature Field A Include Plots Back User ADM Status pos Syros 5 Click Back to save changes Editing Favorites You can add assays and select the order in which they appear on the main menu using the Edit Favorites option All user levels can add and organize assays in the Favorites list 1 Select Setup from the main menu 2 Select Edit Favorites from the Setup menu 3 Select a slot either with a current assay to remove the assay or an empty slot to add an assay BI NOTE If you want to change the order of the assays first remove the assays then add them in the order you want them to appear 78 Muse Cell Analyzer User s Guide Home gt Setup Edit Favorites Favorites
76. ystem Check Beads Refer to the Muse System Check Kit User s Guide for information 2 Select System Check under Essential Tools at the main menu to display the System Check screen 3 A message appears prompting you to check the fluid levels in the cleaning and waste bottles Check the fluids then click Close Always remember to reset the fluid levels when you fill the cleaning solution bottle and empty the waste bottle For information on resetting the fluid levels see Resetting Fluid Levels on page 63 System Check 37 4 Select Run System Check Run System Check B 1 View the result of the last EEEN System Check View Log E Last Check 25 MAY 2011 08 26 45 User KVT Status Ready for Check 22 A 10 5 The first time you run the procedure enter the bead lot number expiration date and check code e Enter the Bead Lot and press Done on the keypad e Touch the calendar icon in the Exp Date field to select the expiration date Touch outside the calendar to close it e Touch the Check Code field and enter the code All values are required and can be found on the information card that comes with the bead kit Once you enter this information it will remain in the software Each time you run the procedure check the information to ensure it is accurate Update the values when a new lot number of System Check Beads is used if necessary 38 Muse Cell Analyzer User s Guide 6 Select Nex

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