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Agilent Technologies NovoCyte Quanteon Operator'S Manual Download Page 88

Running Samples 

88  

NovoCyte Quanteon Flow Cytometer Operator's Guide 

compensation matrix. Right-click 

Compensation

 under the 

Compensation 

Specimen

 node in the 

Experiment Manager 

and select 

Copy

, as shown in Figure 

56, then right-click the experimental sample in the

 Experiment Manager 

and select 

Paste

. The second method is to treat the compensation matrix as a template. 

Select

 Compensation

 under the 

Compensation Specimen

 node, drag and drop it 

to the experimental sample to which you want to apply the compensation matrix. 
For detailed information about the template function, please refer to the 

NovoExpress Software Guide

. 

 

NOTE 

You can also use the same method to apply the compensation matrix to a 
specimen. In this case, the same compensation matrix will be applied to all 
samples belonging to the specimen. 

 

Once the compensation matrix is applied to the experimental sample, double-
check the data after compensation and confirm the correctness of the 
compensation.  

 

NOTE 

The compensation matrix generated by automatic compensation cannot be 
modified. However, after it is applied to the experimental samples, you can adjust 
the compensation matrix or spillover matrix if needed. 

 

 

Figure 56.

 

Copy/Paste the Compensation Matrix 

Summary of Contents for NovoCyte Quanteon

Page 1: ...NovoCyte Quanteon Flow Cytometer Operator s Guide For Research Use Only Not for use in diagnostic procedures...

Page 2: ...e material in this document that conflict with these terms the warranty terms in the separate agreement shall control Technology Licenses The hardware and or software described in this document are fu...

Page 3: ...rmation on how to install NovoCyte Quanteon instrument and associated standard accessories including installation warnings space and environment requirements instructions for lifting and carrying and...

Page 4: ...History Appendix A NovoCyte Quanteon Laser and Detection Channel Configuration This chapter provides additional information on NovoCyte Quanteon laser and detection channel configuration Appendix B No...

Page 5: ...0 Laser Product Classification 21 Biosafety 21 Limitations 22 Agilent Regulatory Compliance Statement 23 CE Compliance 23 Electromagnetic Compatibility 23 EMC Declaration for South Korea 25 Detachable...

Page 6: ...ument Reagents 44 QC Particles 44 System Components 44 NovoCyte Quanteon Instrument 45 NovoCyte Workstation and Monitor 46 NovoCyte Fluidics Station II 47 NovoCyte Fluidics Cart Optional 49 NovoSample...

Page 7: ...tion 58 Add Instrument Reagents 59 Empty Waste 60 Verify and Modify Instrument Configuration 61 View and Verify Instrument Configuration 61 Modify Instrument Configuration 61 Instrument QC Test 68 Ins...

Page 8: ...id Ultrafiltration UF Filter Optional 122 Unscheduled Maintenance 125 Debubble 125 Clean the External Surface of the Instrument 126 Decontamination 126 Prime Fluidic System before Long Term Storage 12...

Page 9: ...s Guide 9 Detection Channel Configuration 139 Appendix B NovoCyte Quanteon Technical Specifications 143 NovoCyte Quanteon Technical Specifications 143 NovoCyte Workstation and Monitor Specifications 1...

Page 10: ...Contents NovoCyte Quanteon Flow Cytometer Operator s Guide 10 This page is intentionally left blank...

Page 11: ...please refer to NovoExpress Software Guide For additional information about NovoSampler Q please refer to NovoSampler Q Operator s Guide For additional information about NovoCyte Fluidics Cart please...

Page 12: ...ity with the provisions of relevant directives for CE marking SGS marking This symbol indicates SGS has evaluated the safety of this product to comply with the provisions of relevant standards for US...

Page 13: ...isturbances COM COM communication port USB USB communication port NET Network communication port The following precaution labels appear on Agilent flow cytometer and associated accessories to indicate...

Page 14: ...s not allowed to remove or disassemble the parts Doing so will void the warranty on the product Glossary The following table lists the glossary which may appear in this guide Table 4 Glossary used in...

Page 15: ...Dye PD Photodiode PE Phycoerythrin Dye PE Alexa Fluor Phycoerythrin and Alexa Fluor Tandem Dye PE Cy5 Phycoerythrin and Cyanine 5 Tandem Dye PE Cy5 5 Phycoerythrin and Cyanine 5 5 Tandem Dye PE Cy7 Ph...

Page 16: ...Ltd Pacific Blue and Pacific Orange are trademarks of Life Technologies Corporation Alexa Fluor and Texas Red are registered trademarks of Life Technologies Corporation Qdot is a registered trademark...

Page 17: ...wer verify that all safety precautions are taken according to the operator s guide of the individual instrument Verify that the voltage range and frequency of your power distribution matches to the po...

Page 18: ...ration until they can be repaired by qualified service personnel WARNING The handling of toxic and hazardous solvents and flammable liquids can hold health risks When working with solvents ensure to o...

Page 19: ...imal operation of the instrument WARNING The sample injection probe is a mechanical movable part and may pinch or injure your fingers To prevent accidents do not put your hand underneath the sample in...

Page 20: ...ment for about 6 7 seconds Immediately contact your local Agilent Technologies representative for support If the power cord becomes frayed broken or damaged please contact Agilent Technologies Do not...

Page 21: ...aintenance and repair can only be carried out by qualified Agilent service professionals WARNING In order to avoid the risk of exposure to laser light be sure to follow the guidelines listed below Mod...

Page 22: ...solution and waste may contain materials controlled by contamination regulations and emission standards Please follow local regulations and guidelines for proper disposal Dispose of waste in accordanc...

Page 23: ...ompatibility EMC Directive Low Voltage Directive LVD Machinery Directive MD RoHS Directive etc Agilent has confirmed that each product complies with the relevant Directives by testing samples against...

Page 24: ...tibility in other environments due to conducted as well as radiated disturbances If this equipment does cause harmful interference to radio or television reception which can be determined by turning t...

Page 25: ...ust only use the power cord that was shipped with this product Do not use this power cord with any other product Sound Emission Certification for Federal Republic of Germany Sound pressure Sound press...

Page 26: ...ase refer to the sections relating to the instrument operation in this guide If a problem occurs refer to Troubleshooting in this guide for solutions For additional technical support please contact yo...

Page 27: ...chnical Support Request in the NovoExpress main window The Technical Support Request Creation Wizard automatically collects the instrument configurations system logs current screenshot current experim...

Page 28: ...Prologue 28 NovoCyte Quanteon Flow Cytometer Operator s Guide This page is intentionally left blank...

Page 29: ...ell cycle analysis of the nucleus analysis of cell proliferation and GFP reporter gene detection etc Agilent flow cytometer can detect a particle with the size of 0 1 m 50 m Besides human cells it can...

Page 30: ...l other than the standard configuration is available Please refer to Appendix A in this guide for more details of your product Please contact Agilent local sales representatives or distributors for cu...

Page 31: ...ent light 2 whereas the reflected light is reflected in a 180 angle relative to the incident light 1 and 3 Figure 2 Figure 2 Bandpass Filter NOTE Refer to Appendix A in this guide for list of the opti...

Page 32: ...Shortpass filters SP pass light with a shorter wavelength than the cutoff wavelength For example a 472 nm shortpass filter allows wavelengths shorter than 472 nm to pass through it and either absorbs...

Page 33: ...lar range to be detected by the detector Commonly the name of the bandpass filter is used interchangeably with fluorescence channel to specify the detection wavelength range For example PE is routinel...

Page 34: ...ir ground state shortly accompanied by the release of energy Most energy is released in the form of emitted light whereas a very small portion of the energy is released in the form of heat The light e...

Page 35: ...two or more fluorochromes are used in the same test the overlap in wavelength ranges often makes it impossible to isolate light from a given fluorochrome from another fluorochrome using optical filte...

Page 36: ...C positive population downwards until it has the same Mean or Median value of PE signal as the unstained population Figure 10 Figure 9 Theoretical Display of FITC vs PE without Compensation Figure 10...

Page 37: ...ions Users can modify the compensation matrix manually or use the automatic compensation function Fluorescence compensation can be conducted during or after sample acquisition NOTE For more informatio...

Page 38: ...the SIP after each sample acquisition The waste fluid generated by SIP cleaning procedure is collected to the waste container This automatic SIP cleaning function ensures minimal inter sample carryov...

Page 39: ...er uses a high precision syringe pump to aspirate the sample and drive it into the flow cell for analysis This sampling method can precisely control the volume of the sample for accurate volumetric co...

Page 40: ...important for obtaining good quality data A higher sample flow rate is generally used for qualitative measurements such as immunophenotyping The data is less resolved but is acquired more quickly A l...

Page 41: ...atter or fluorescence emission has a very brief duration usually only a few microseconds This brief emission of light is converted into an electrical signal a pulse shown in Figure 14 by the photodete...

Page 42: ...nd every selected fluorescence channel Width is based on the FSC signal One effective event has one Width parameter Pulse Digitization The electronics system digitizes the pulse produced by the photod...

Page 43: ...NovoCyte Quanteon Flow Cytometer Operator s Guide 43 Figure 16 Pulse Digitization System Requirements Software NovoExpress software Workstation NovoCyte Workstation Fluidics Station NovoCyte Fluidics...

Page 44: ...oFlow Sheath Fluid NovoClean Solution NovoRinse Solution NOTE Please consult the Agilent website or contact your local representative for more detailed ordering information QC Particles Agilent Flow C...

Page 45: ...Flow Cytometer NovoCyte Quanteon Instrument The instrument front side Figure 18 has a power switch an LED status indicator LED light and a front panel cover By default a single tube holder is pre inst...

Page 46: ...ication ports and five fluidic connection ports The power port is used to supply the power to the instrument through provided power cord The communication ports are used to communicate the instrument...

Page 47: ...low sheath fluid container 3 L one NovoRinse solution container 500 mL one NovoClean solution container 500 mL and one waste container 3 L by default Figure 21 The four containers should be placed in...

Page 48: ...status indicator on the front of the NovoCyte Fluidics Station II also give information on the fluid level and are summarized in the following table Table 6 NovoCyte Fluidics Station II LED Status In...

Page 49: ...ovoCyte Fluidics Cart Optional Agilent flow cytometer can also be connected to optional NovoCyte Fluidics Cart with larger NovoFlow and Waste volume capacity The NovoCyte Fluidics Cart comes with one...

Page 50: ...n 50 NovoCyte Quanteon Flow Cytometer Operator s Guide please visit www agilent com or contact Agilent sales representative For details about the NovoSampler Q please refer to the NovoSampler Q Operat...

Page 51: ...o the workstation through the USB cable before installing or relocating the instrument WARNING Do not operate the instrument in an environment where potentially damaging liquids or gas are present CAU...

Page 52: ...g lifetime or cause damage to the instruments Instruction for Lifting and Carrying Do not attempt to lift or move the instrument without the assistance of others Improper lifting can cause painful and...

Page 53: ...Install NovoCyte Quanteon A complete and typical installation of NovoCyte Quanteon instrument requires installation of instrument workstation monitor and NovoCyte Fluidics Station II When NovoSampler...

Page 54: ...he NovoCyte workstation Connection D in Figure 23 NOTE There are four communication ports at the back of NovoCyte Quanteon instrument see Figure 24 COM 1 port connects to the NovoSampler Q optional CO...

Page 55: ...ntamination solution and is connected by the yellow tubing Waste 1 and Waste 2 connect to the red tubing Figure 25 Interface of the Fluidics Ports The reagent containers are connected through the same...

Page 56: ...0 240 VAC 50 60Hz by connecting one end of the supplied power cord to the power port on the instrument Connection G in Figure 23 and connect the other end of the power cord to the power supply WARNING...

Page 57: ...connection between the NovoCyte Workstation and the instrument are correct and secure 4 Check the NovoFlow sheath fluid container the NovoRinse solution container and the NovoClean solution container...

Page 58: ...the status bar which indicates that the instrument is properly connected and that the instrument is in the initialization process The initialization process takes several minutes After completion the...

Page 59: ...ect the quick coupler on the cap and gently place the container onto the NovoCyte Fluidics Station II Be careful to prevent any twisting or kinking of the tubing 6 Ensure the instrument is in Ready st...

Page 60: ...ore details 1 Make sure the instrument is in Ready status or the power is turned off 2 Disconnect the tubing from the waste container by gently pressing the metal clip of the quick coupler on the cap...

Page 61: ...uration users need to replace the appropriate optical filters of the system following the prompt in the Instrument Configuration window To make the new optical configuration effective a QC Test with a...

Page 62: ...the configuration if needed Click Change Optical Configuration to continue NOTE A customized optical configuration can be deleted by clicking Delete Only the user defined optical configurations can b...

Page 63: ...e 63 Figure 29 Change the Optical Filter or Dichroic Mirror 4 Click Complete The software will automatically read and check the validity of the new optical configuration Figure 30 Complete Modifying t...

Page 64: ...low Cytometer Operator s Guide Figure 31 Confirm the New Optical Configuration NOTE If the new optical configuration is invalid there will be message in Instrument Configuration window Figure 32 Dialo...

Page 65: ...g the NovoExpress 7 After NovoExpress is restarted the following window will appear Click QC Test to continue Figure 34 Dialog Window for Performing the QC Test 8 Properly prepare 1 mL QC particles sa...

Page 66: ...e will automatically adjust the photodetector gain by running the QC particles When the adjustment is completed the software will automatically perform the QC Test Click Report when the QC test is com...

Page 67: ...s below make sure the QC particles are properly prepared Run the QC particles once again after the correct action has been taken Ensure there is at least 400 L sample remaining in the sample tube Clic...

Page 68: ...e installation and on a daily basis to ensure the consistency of the instrument performance Please refer to NovoExpress Software Guide for detailed instructions Instrument Shutdown At the end of the d...

Page 69: ...h light goes out NOTE When shutdown is initiated from NovoExpress software if no additional function option is selected the instrument automatically performs the shutdown cleaning process and turns of...

Page 70: ...Instrument Operation 70 NovoCyte Quanteon Flow Cytometer Operator s Guide This page is intentionally left blank...

Page 71: ...OTE The fluorescence channel configuration varies for different instrument models Please refer to Appendix A in this guide for optical configuration of your product This example will create a specimen...

Page 72: ...on Cytometer Control panel to create a new sample to start the experiment Figure 40 shows the NovoExpress software interface The status bar indicates that the instrument is properly connected and is...

Page 73: ...need to be created Repeat creating a new sample to add Sample 2 to the same specimen Rename specimen and sample name by click the name twice or select Rename on the right click drop down menu In this...

Page 74: ...nd Sample Define Sample Acquisition Parameters 1 Double click the T Lym sample to make it the Active Sample 2 Set the test parameters in the Cytometer Setting Parameters panel see Figure 42 By default...

Page 75: ...e available for data analysis However selecting too many parameters other than those really needed in the experiment will unnecessarily increase the size of the experiment file causing decreased data...

Page 76: ...on uses fixed photodetector gain by default Adjustable photodetector gain is available as advanced function Please refer to the NovoExpress Software Guide for details about photodetector gain adjustme...

Page 77: ...ed such as the size of the cells the resolution to be achieved the concentration of the sample the throughput etc In general it is advisable to choose a core diameter that is similar or slightly large...

Page 78: ...w value when clicking the desired position on the plot for the subsequent data acquisition NOTE For a new experiment it might be difficult to determine how to set the threshold In this case conduct a...

Page 79: ...000 1 000 000 Use linear or logarithmic scale for FSC and SSC Cells lines smaller than 20 m in cell diameter 100 000 300 000 Use linear or logarithmic scale for FSC and SSC Fixed or un fixed freshly...

Page 80: ...le At the same time the sample name in Cytometer Control Active Sample Information panel becomes T Lym To begin collecting this sample place the CD3 FITC CD8 PE CD45 PerCP CD4 APC sample tube into the...

Page 81: ...48 Active Sample Information Once the stop condition is reached sample acquisition will automatically stop NOTE You can also manually stop the sample acquisition by clicking the Stop button in the Cy...

Page 82: ...worn when handling the waste which should be disposed of in accordance with local safety regulations Perform Fluorescence Compensation Fluorescent molecules have their own spectral emission ranges and...

Page 83: ...sample can be generated from the same biological sample as the experimental sample or compensation beads may be used Compensation beads are microspheres with capturing agent on the surface The capturi...

Page 84: ...dian as Parameter for Calculation Figure 50 Click the OK button and a new Compensation Specimen will appear in the Experiment Manager under the experiment file node This Compensation Specimen contains...

Page 85: ...lts You can also change the settings by right clicking the Compensation Specimen in the Experiment Manager Figure 51 and select Auto Compensation Setup Figure 51 Compensation Specimen and Related Sing...

Page 86: ...own in Figure 52 Figure 52 Data Analysis for PE Single Stained Sample in Auto Compensation Under normal circumstances the automatically generated gates are appropriate However due to the complexity of...

Page 87: ...Specimen node in the Experiment Manager Figure 53 Double click Compensation to View the Compensation Matrix Figure 54 Compensation Matrix Generated by Auto Compensation Figure 55 Spillover Matrix Gene...

Page 88: ...ation matrix For detailed information about the template function please refer to the NovoExpress Software Guide NOTE You can also use the same method to apply the compensation matrix to a specimen In...

Page 89: ...ation to open the New Auto Compensation window Select Import Samples from FCS files NOTE This window can also be accessed by right clicking the experiment file and selecting New Auto Compensation in t...

Page 90: ...tware Click OK to continue Figure 59 Dialog Box for Successful Importing FCS Files 4 Set the compensation parameters in the New Auto Compensation window i e Area or Height Median or Mean Ensure the co...

Page 91: ...OTE The software will automatically associate the imported FCS file with the selected channel parameter based on the keyword of the file name e g B525 Users need to verify and select the correct FCS f...

Page 92: ...ain the sample as the X axis parameter and each of the other fluorescence channels as the Y axis parameter Apply the main population gate to all the fluorescence channel density plots For example in t...

Page 93: ...ation in the bottom right segment Use the quick compensation sliding bar or adjust the coefficient in the spillover matrix to conduct the compensation The correct compensation is achieved when the Mea...

Page 94: ...PerCP and APC Channels NOTE Select Preview in the compensation window to visualize the compensation results dynamically when adjusting the spillover matrix coefficient Click the Expand Statistics icon...

Page 95: ...over Matrix for PerCP Single Stained Sample Figure 66 Spillover Matrix for APC Single Stained Sample Apply the Compensation Matrix To use the compensation coefficient generated by the single stained s...

Page 96: ...ts of the T Lym sample before and after the compensation respectively Figure 67 Incorporate the Spillover Coefficients from Single Stained Samples into the Spillover Matrix Figure 68 Results of T Lym...

Page 97: ...to biexponential scale to have a better visualization of the data Analyze Data The NovoExpress software provides a variety of plots and gates for flow cytometry data analysis The type of plots include...

Page 98: ...Sample in Data Acquisition Green Arrow Create Plots and Gates Firstly we need to look at the CD45 PerCP vs SSC plot to locate the target cell population i e the lymphocytes which have relatively large...

Page 99: ...the mouse button the range of the plot is automatically enlarged according to the zoom in operation You can also Undo the Zoom In operation by clicking the undo icon or click the zoom out icon from th...

Page 100: ...ate the Sample Use the method described in Perform Fluorescence Compensation in this guide to apply the compensation matrix created by either automatic compensation or manual compensation to the sampl...

Page 101: ...ady include data Newly acquired data will be appended to the existing data for the sample 1 Select the sample to be the Active Sample Set the Stop Condition for the acquisition 2 Load the sample into...

Page 102: ...only delete events Outside or Inside a specified gate Figure 76 Delete Events Save Data The NovoExpress software automatically saves the data along with the cytometer settings information to the expe...

Page 103: ...and select Export Export FCS File or Export Export CSV File as shown in Figure 77 In the prompt window specify the folder to export the data Figure 77 Manually Export Data The data can be automatical...

Page 104: ...Running Samples 104 NovoCyte Quanteon Flow Cytometer Operator s Guide Figure 78 Enable the Automatic Data Exporting Function Figure 79 Configure Exporting Settings...

Page 105: ...men in the Experiment Manager right click and select Import Import FSC File as shown in Figure 80 Figure 80 Import FCS File NOTE The NovoExpress software provides powerful data analysis functions such...

Page 106: ...Running Samples 106 NovoCyte Quanteon Flow Cytometer Operator s Guide This page is intentionally left blank...

Page 107: ...ng manual maintenances Scheduled Maintenance The following maintenance should be conducted on regular basis Table 9 Scheduled Maintenance and Recommended Frequency Maintenance Recommended Maintenance...

Page 108: ...ocedure below to manually clean the outer surface of the sample injection probe NOTE The procedures described below may vary if NovoSampler Q is used Please refer to NovoSampler Q Operator s Guide for...

Page 109: ...NING Sample injection probe will move downward after clicking the OK button Do not put your fingers underneath the sample injection probe 5 Prepare a cotton swab or a damp soft cloth with NovoClean so...

Page 110: ...UTION Sample injection probe is fragile and may be damaged if handled improperly When cleaning the sample injection probe please be careful not to deform or bend it 6 Click OK button after cleaning Fi...

Page 111: ...Turn off the power of the instrument 2 Prepare a cotton swab soaked with NovoClean solution 1X Wipe the SIP cleaning apparatus lower surface back and forth until it is clean Figure 86 Wipe the SIP Cl...

Page 112: ...NovoExpress Sterilize Instrument Reagent Containers It is very important to keep the instrument reagent free of contamination by sterilizing the instrument reagent containers This procedure is highly...

Page 113: ...m filtered DI water or NovoFlow sheath fluid Soak for 20 min and empty the reagent container 6 Wipe the outer surface of the container with a dry and clean soft cloth 7 Place the container back onto t...

Page 114: ...any time by clicking the Instrument Replace Fluidic System Consumables icon in the NovoExpress software when the instrument is in Ready status Figure 90 Replace Fluidic System Consumables icon Click...

Page 115: ...on to continue If Cancel is selected a dialog box below will be prompted The process can be aborted by clicking Yes button Figure 92 Software Message for Cancelling Fluidics System Consumable Replacem...

Page 116: ...em Consumables During the Priming procedure the progress of the Priming will be shown below in the NovoExpress software Figure 95 Software Message Indicating the Progress of Priming NOTE The Priming p...

Page 117: ...cement The following sections describe the detailed procedure for replacing fluidic system consumables It is highly recommended to replace all the fluidic system consumables at the same time to ensure...

Page 118: ...uer connectors only to avoid damage of the filters and filter leakage 5 Once the instrument is Ready click Instrument Fluidics Maintenance Priming to prime the fluidic system Observe the sheath fluid...

Page 119: ...te Filter 1 Make sure the instrument is in Ready or shut down status 2 Open the front panel of the instrument Locate the waste filter as shown below Figure 98 Location of Waste Filter 3 Unscrew the Lu...

Page 120: ...e is no leakage through the filter tubing connections Then run the Priming process when start up process is completed and instrument is ready CAUTION When replacing the waste filter do not twist or ki...

Page 121: ...se use proper force when tightening the Luer connectors CAUTION The ends of the sheath fluid inlet filter are different to ensure the filter is connected into the system in the correct direction Incor...

Page 122: ...s recommended to replace this filter every two weeks after the first time use by following the procedures below These procedures can also be used when installing this filter is needed NOTE Agilent flo...

Page 123: ...et of the new filter to the female Luer connector and the inlet of the new filter to the male Luer connector Secure the filter onto the filter rack by gently sliding it through the open slit on the ra...

Page 124: ...dic system Ensure no fluidic leakage from the filter s inlet outlet 6 Load a sample tube with 1 mL Milli Q or 0 2 m filtered DI water onto the tube holder Create a new experiment Set Stop Condition as...

Page 125: ...uspecting contamination in the system Prime Fluidics System before Long Term Storage No plan to run the instrument for more than two weeks Prime Fluidics System after Long Term Storage Instrument has...

Page 126: ...y biological samples which may be biohazardous Ensure to wear appropriate personal protective equipment e g gloves clothing and eyewear when cleaning the surface CAUTION Do not use isopropyl alcohol I...

Page 127: ...etion Figure 106 NovoExpress Dialog Box to Guide the Decontamination Procedure NOTE Contact your local Agilent representative or distributor for any questions regarding this procedure Prime Fluidic Sy...

Page 128: ...ng process is also recommended before using the instrument to ensure the fluidic system is working in optimal condition 1 Sterilize the reagent containers following the procedure described in Steriliz...

Page 129: ...the purging procedure Follow the instructions in each step and click OK to move to the next step until completion Figure 109 Software Message When Starting the Purge Procedure 3 Power off the instrume...

Page 130: ...ly available when Calibrate Fluidics Station privilege is enabled Refer to NovoExpress Software Guide for instructions to enable this privilege 1 Click Instrument Operation Calibrate Fluidics Station...

Page 131: ...of optical filter Gently hold the proper optical filter and pull it upward to remove it from the slot Figure 29 2 Blow off the contaminants on the surface of optical filters Many contaminants are just...

Page 132: ...or every new cleaning attempt Clean Fluidic System When the CV of the data is larger than normal or the QC Test fails or excessive debris appears it is possible that the flow cell or sampling tubing i...

Page 133: ...isted in these two tables NOTE Investigate possible causes and perform the recommended solutions in the given order Table 11 Instrument Troubleshooting Guide Observation Possible Causes Recommended So...

Page 134: ...ubble function from NovoExpress Software Flow cell is clogged Execute the Unclog function from NovoExpress software Improper sample preparation Re prepare the sample properly The sample flow rate is s...

Page 135: ...ation Make sure the plate or tube rack is fully seated onto the sample tray properly NovoSampler or NovoSampler Pro or NovoSampler Q is not calibrated Re calibrate the NovoSampler or NovoSampler Pro o...

Page 136: ...Troubleshooting 136 NovoCyte Quanteon Flow Cytometer Operator s Guide This page is intentionally left blank...

Page 137: ...NovoCyte Quanteon Flow Cytometer Operator s Guide 137 8 Version History Table 12 History of updates Date Version Changed by Description 08 31 2018 Initial Release 10 31 2019 Second Edition 03 31 2021...

Page 138: ...Version History 138 NovoCyte Quanteon Flow Cytometer Operator s Guide This page is intentionally left blank...

Page 139: ...available Please contact your local Agilent Biosciences representatives or distributors for more details Table 13 lists the typical detectable fluorochromes on NovoCyte Quanteon standard systems Tabl...

Page 140: ...685 LP 660 20 PE Cy5 Y667 667 30 Y695 705 LP 695 40 PE Cy5 5 Y725 FRM 725 40 PE Alexa Fluor 700 Y780 757 LP 780 60 PE Cy7 405 nm V445 495 LP 445 45 Pacific Blue V525 552 LP 525 45 AmCyan V530 530 30 V...

Page 141: ...anteon System1 637 nm R660 685 LP 660 20 APC R667 667 30 R695 705 LP 695 40 Alexa Fluor 680 R725 FRM 725 40 Alexa Fluor 700 R780 757 LP 780 60 APC Cy7 1 Certain fluorescence detection channels bandpas...

Page 142: ...NovoCyte Quanteon Laser and Detection Channel Configuration 142 NovoCyte Quanteon Flow Cytometer Operator s Guide This page is intentionally left blank...

Page 143: ...100 mW Laser Beam Profile 10 x 60 m elliptical beam Optical Alignment Procedure Fixed no operator alignment required Fluorescence Detection Silicon Photomultiplier SiPM with high photon detection eff...

Page 144: ...SIP surface after each sampling Fluidics System Monitoring In line pressure sensor monitors the pressure in real time Automated system recovery when pressure is out of range due to clogging Fluidics...

Page 145: ...x 75 mm tube 24 well 48 well 96 well flat V U bottom 384 well microtiter plates Agilent Flow Cytometer 24 tube cooling rack for 12 x 75 mm tube Agilent Flow Cytometer 96 well flat U V bottom plate co...

Page 146: ...igh resolution flat panel with resolution of 1 920 x 1 080 or above Computer Operating System Microsoft Windows 10 Professional 64 bit or above with Microsoft Office and NovoExpress software pre insta...

Page 147: ......

Page 148: ...ent Operation Running Samples System Maintenance Troubleshooting Version History Appendix A NovoCyte Quanteon Laser and Detection Channel Configuration Appendix B NovoCyte Quanteon Technical Specifica...

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