Maintenance Guide - ACEA Biosciences Inc. · This maintenance guide describes the maintenance of...

Maintenance Guide

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NovoCyte® Flow Cytometer

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Table of Content

Prologue 5

I. Revision History ........................................................................................................................5

II. Contact Information .................................................................................................................6

III. Trademark ...................................................................................................................................6

IV. Using this Guide ........................................................................................................................6

V. Conventions ...............................................................................................................................6

VI. Technical Support .....................................................................................................................7

1. Scheduled Maintenance 9

1.1 Preventative Maintenance ................................................................................................... 10

1.2 Replacing Fluidic System Consumables ........................................................................... 14

2. Unscheduled Maintenance 21

2.1 Remove Bubbles .................................................................................................................... 21

2.2 Unclog the Sample Injection Probe (SIP) ........................................................................ 22

2.3 Clean the External Surface of the Instrument ................................................................. 23

2.4 Clean the Sample Injection Probe (SIP) ........................................................................... 24

2.5 Decontamination ................................................................................................................... 26

2.6 Clean Fluidic System before Long-Term Storage ............................................................ 27

2.7 Purge Fluidic System before Shipment ............................................................................. 28

2.8 Calibrate the Fluidics Station .............................................................................................. 29

2.9 Clean the Optical Filters and Mirrors ................................................................................ 30

3. Troubleshooting 31

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Table of Content

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Prologue

Revision History

Prologue

I. Revision History

Version Revision Date

1.0 2015.12

1.1 2016.08

1.2 2017.05

1.3 2018.08

© Copyright 2018, ACEA Biosciences, Inc. All rights reserved. The product description and its corresponding intellectual property rights belong to ACEA Biosciences, Inc.

The content in this guide may not be reproduced in any form, transmitted, transcribed, stored in a retrieval system, or translated into any language or computer language, in any form or by any means without prior written permission from ACEA Biosciences, Inc.

ACEA NovoCyte® © ACEA Biosciences, Inc. ACEA NovoCyte® flow cytometer and re-lated reagents are the property of ACEA Biosciences, Inc.

ACEA NovoExpress® © ACEA Biosciences, Inc. ACEA NovoExpress® software is the property of ACEA Biosciences, Inc. Each sale of a stored unit of this soft-ware grants the purchaser a nonexclusive personal license. This software may not be duplicated, reproduced, or copied in any form or by any means whatsoever, except as otherwise permitted by law.

The information in this guide is subject to change without notice. ACEA Biosciences reserves the right to change its products and services at any time to incorporate the lat-est technological developments. Even though this guide has been prepared with every precaution to ensure accuracy, ACEA Biosciences assumes no liability for any errors or omissions, nor for any damages resulting from the application or use of this information. ACEA Biosciences welcomes customer input on corrections and suggestions for improve-ment.

Questions or comments regarding the contents of this guide can be directed to the address below or to your local ACEA Biosciences representative.

ACEA Biosciences, Inc. 6779 Mesa Ridge Road, Suite 100 San Diego, CA 92121 USA

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Prologue

Contact Information

II. Contact Information

Manufacturer ACEA Biosciences, Inc.6779 Mesa Ridge Road, Suite 100San Diego, CA 92121USA

China Distributor ACEA Bio (Hangzhou) Co., Ltd. Building 5, 2# Xiyuan Wu Rd., Xihu DistrictHangzhou, Zhejiang Province, P. R. China310030

International Distributors Please contact ACEA Biosciences, Inc., or visit www.aceabio.com for a complete list of interna-tional distributors

III. TrademarkACEA Biosciences, the ACEA logo, NovoCyte®, NovoExpress®, NovoSampler®, Novo-Flow®, NovoRinse®, and NovoClean®, are trademarks of ACEA Biosciences, Inc., in the United States and some other countries.

All other company and product names might be trademarks of the respective companies with which they are associated.

IV. Using this GuideThis maintenance guide describes the maintenance of the ACEA NovoCyte flow cytom-eter. Most of these functions are controlled by the ACEA NovoExpress software. For ad-ditional information about the ACEA NovoCyte, please refer to NovoCyte® Flow Cytom-eter Operator’s Guide. For additional information about the ACEA NovoExpress software, please refer to NovoExpress® Software Guide.

ACEA NovoCyte Flow Cytometer Maintenance Guide assumes that the user is familiar with the Microsoft Windows® operating system. For questions regarding the Microsoft Windows® operating system, see the operating system’s help and support files.

V. Conventions

Text Conventions

To impart information that is consistent and easy-to-read, the following text conventions are used in this guide:

Format Description

Numbered Listing 1 2 ......

Describes the steps in a procedure that must be performed in the listed order.

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Prologue

Technical Support

Format Description

Italic font, gold Points to a different chapter in this guide, which should be referred to for better understanding.

Italic font Describes buttons, icons or functions when operating the NovoExpress software. In addition, important notes and in-formation are also shown in italic font.

→ Indicates the sequence of the menu operation in NovoEx-press software. For example, File → Print means to select the Print function from the File menu.

Ctrl+X When used with keyboard characters, + means to press two keys simultaneously. For example, Ctrl+C means to hold down the Control key while pressing the letter C key.

Symbols

The following table lists the symbols used in this guide:

Symbol Meaning Description

Biohazard This symbol indicates that certain precautions must be taken when working with potentially infectious bio-logical specimens and materials.

Laser Radiation This symbol indicates that the operation could po-tentially cause exposure to laser radiation. Protective measures should be taken during the operation.

Warning This symbol indicates that improper or unsafe opera-tion could result in instrument damage, data loss, and personal injury.

Shock Hazard This symbol indicates that the operation has a risk of electric shock.

Important Note This symbol indicates information which is critical to the success of the procedure or use of the product.

Additional Informa-tion

This symbol provides additional information about the current topic or procedure.

uuu Form Table continues on next page.

n The End of the Table End of a table.

VI. Technical SupportIf there are operating or technical questions, please refer to the sections relating to the in-strument operation in this guide. For additional technical support, please contact your lo-cal ACEA Biosciences representative or distributor. When contacting ACEA Biosciences, be sure to provide the following information:

► The product name, product model and serial number.

► Usage history of the instrument.

► Instrument status information on the status bar from the NovoExpress software. Or if there is a warning or error message, please also provide that information.

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Prologue

Technical Support

► Experiment information conducted on the instrument, if not confidential.

► Details of recent instrument QC test.

For support within the US, please call 866-308-2232.

For support within China, please call 400-600-1063.

For users in other countries or regions, contact your local ACEA Biosciences representa-tives or distributors, which may be found at www.aceabio.com.

When encountering an issue during operation of NovoCyte flow cytometer, NovoSam-pler or NovoSampler Pro instrument, it is highly recommended to submit a Technical Support Request by clicking Home → Technical Support Request in the NovoExpress main window. The Technical Support Request Creation Wizard automatically collects NovoCyte configurations, NovoExpress system logs, current screenshot, current experi-ment file and other information that helps in the diagnosis and troubleshooting of No-voCyte instrument. User can also attach any other files using this function. Please refer to the NovoExpress® Software Guide for detailed information on this function.

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Scheduled Maintenance

1. Scheduled Maintenance

It is important to conduct instrument maintenance on a regular basis in order to achieve optimal performance for the instrument and obtain reliable data. ACEA NovoCyte incor-porates automatic procedures for maintenances as much as possible, eliminating time-consuming manual maintenance. This chapter describes the procedures to execute or con-duct the maintenance procedures.

Table 1-1 lists the regular maintenance to be done on the NovoCyte instrument.

Maintenance Recommended Maintenance Frequency

Daily Start-up Daily. Automatic system cleaning during start-up.

Daily Shutdown Daily. Automatic system cleaning and decontam-ination during shutdown.

Add Instrument Reagents (ACEA No-voFlow Sheath Fluid, ACEA NovoRinse Solution, and ACEA NovoClean Solu-tion)

As needed. Check the instrument reagent volume in the containers before each experiment and add more reagents when necessary.

Empty Waste As needed. Check waste volume before each ex-periment and empty waste container when nec-essary.

Preventative Maintenance

► Clean the Sample Injection Probe (SIP)

► Clean the SIP Cleaning Apparatus

► Conduct Extensive Rinse

► Unclog the Flow Cell

► Sterilize Instrument Reagent Containers

Every month

Replacing Fluidic System Consumables

► Replace the ACEA Sheath Fluid In-Line Filter

► Replace the Waste Filter

► Replace the ACEA NovoFlow Sheath Fluid Inlet Filter

► Replace the ACEA NovoRinse and ACEA NovoClean Solution Inlet Filters

Every two months or prompted by NovoExpress Software

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Table 1-1 Scheduled Maintenance and Recommended Frequency

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1.1 Preventative Maintenance

It is recommended to conduct preventative maintenance once every month to ensure the fluidic system is in good condition.

► Clean the Sample Injection Probe (SIP)

Materials

► Clean soft cloth

► ACEA NovoClean solution (1X)

► Deionized (DI) water

Procedure

1 Turn off the power of the instrument.

2 Open the top cover of the instrument.

3 Locate the SIP.

Figure 1-1 Locate the SIP

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4 Unscrew the brown connector by turning it counter clockwise.

Figure 1-2 Unscrew the Brown Connector

Sample injection probe directly contacts the biological samples which may be biohazardous. Wear gloves when handling the sample injection probe.

Pay attention to the ferrule (indicated with a red arrow in Figure 1-3) at the end of the tubing after removing the brown connector from the slot. Handle the as-sembly carefully and do not remove the ferrule from the tubing. Place the entire assembly on a clean lint-free wipe.

Figure 1-3 Ferrule at the End of Tubing after Removing the Connector

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5 Remove the SIP straight up from the housing slot.

Figure 1-4 Take SIP out from the Housing Slot


Sample injection probe is fragile and may be damaged if handled improperly. When cleaning the sample injection probe, please be careful and prevent deforming or bending it.

6 Prepare a damp cloth with ACEA NovoClean solution (1X). Wipe the sample injection probe’s surface back and forth until the surface is cleaned.

7 Prepare a damp cloth with DI water. Wipe the sample injection probe back and forth for at least five times to clean the ACEA NovoClean solution residue.

8 Put SIP back into the housing slot after cleaning. Make sure that the SIP goes through the hole of the SIP Cleaning Apparatus and the SIP is straight.

Figure 1-5 Put SIP back into the Housing Slot

9 Install the brown connector and tubing by finger tightening it clockwise.

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10 Power on ACEA NovoCyte instrument and start NovoExpress software. After the instru-ment is Ready, click Instrument → Fluidics Maintenance → Backflush to flush the sample injection probe.

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► Clean the SIP Cleaning Apparatus

Materials




Procedure



3 Prepare a damp cloth with ACEA NovoClean solution (1X). Wipe the SIP cleaning ap-paratus back and forth at least five times.

4 Prepare a damp cloth with DI water. Wipe the SIP cleaning apparatus back and forth at least five times.

5 Power on NovoCyte instrument and start NovoExpress software. After the instrument is Ready, click Instrument → Fluidics Maintenance → Backflush to flush the sample injec-tion probe.

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SIP cleaning apparatus directly contacts the biological samples which may be bio-hazardous. Wear gloves when handling the SIP cleaning apparatus.

When cleaning the SIP cleaning apparatus, please be careful to prevent damaging the apparatus and sample injection probe.

► Conduct Extensive Rinse

When NovoCyte is Ready click Instrument → Fluidics Maintenance → Extensive Rinse from NovoExpress software, and the Extensive Rinse procedure will automatically be ex-ecuted. The progress of the procedure will be shown in the NovoExpress status bar.

► Unclog the Flow Cell

When the NovoCyte is in Ready status, click Instrument → Fluidics Maintenance → Un-clog from NovoExpress software, and the Unclog procedure will automatically be execut-ed. The progress of the procedure will be shown in the NovoExpress status bar.

► Sterilize Instrument Reagent Containers

It is very important to keep the instrument reagent free of bacteria for the cytometer. It is highly recommended to clean and sterilize the instrument reagent containers once a month or whenever there is a suspected bacteria growth or possible contamination (e.g. by mishandling the containers). The reagent containers can be cleaned and sterilized using the following method.

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Materials



► Deionized (DI) water (or ACEA NovoFlow solution)

Procedure

1 Wipe the outer surface of the container using medical gauze pads.

2 Empty the reagent container. Fill the reagent container with at least 300 mL of ACEA NovoClean solution (1X) and cap the bottle. Shake the container vigorously to let the ACEA NovoClean solution cover the entire inner surface. Repeat for several times and then soak for 30 min.

3 Empty the reagent container. Fill about 500 mL DI water (or ACEA NovoFlow solution) to reagent container and cap the bottle. Shake reagent container for at least 2 minutes.

4 Repeat step 3 three times.

5 Fill the reagent container all the way to the neck or close to the brim with 0.2 µm filtered DI water or NovoFlow sheath fluid. Soak 20 min and empty the reagent container.

6 Wipe the outer surface of the container with a clean soft cloth. The container is ready to use.

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1.2 Replacing Fluidic System Consumables

The NovoCyte flow cytometer monitors the accumulated running time of the fluidic sys-tem consumables to ensure the consumables are changed in a timely manner for optimal flow cytometry results. When the accumulated running time is reached, the NovoExpress software will prompt a message to remind the user to replace the consumables (Figure 1-6a). Similar message will be prompted when the last replacement is 60 days ago as shown in Figure 1-6b.

Click Replace to continue. Clicking the Close button will cancel the process and allow con-tinued use of the system. However the consumables should be replaced promptly.

Figure 1-6a NovoExpress Software Message for Replacing Fluidic System Consumables When Accu-mulated Running Time is Reached

Figure 1-6b NovoExpress Software Message for Replacing Fluidic System Consumables When Last Replacement is 60 Days Ago

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The fluidic system consumable replacement process can also be initiated manually at any time by clicking the Instrument → Replace Fluidic System Consumables icon in the NovoExpress software when the instrument is in Ready status.

Click the Replace button and a dialog box as shown in Figure 1-7 will be prompted.

Figure 1-7 NovoExpress Software Message for Fluidics Consumables Replacement

The following fluidic system consumables should be replaced at the same time:

► ACEA NovoFlow sheath fluid in-line filter (1pcs) (Cat. No. 2030002)

► Waste filter (1pcs) (Cat. No. 2030003)

► ACEA NovoFlow sheath fluid inlet filter (1pcs) (Cat. No. 2030001)

► ACEA NovoRinse solution inlet filter (1pcs) (Cat. No. 2030005)

► ACEA NovoClean solution inlet filter (1pcs) (Cat. No. 2030007)

Follow the procedure described below to replace all of the fluidic system consumables.

Once all of the fluidic system consumables have been replaced with the new ones, click the Next button to continue.

If Cancel is selected, a dialog box as shown in Figure 1-8 will be prompted. The process can be aborted by clicking Yes button.

Figure 1-8 NovoExpress Software Message for Cancelling Fluidics System Consumable Replacement

A confirmation message will be prompted (Figure 1-9). Click Yes button to inform the system that the consumables have been replaced.

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Figure 1-9 NovoExpress Software Message Confirmation for Fluidics Consumables Replaced

After confirmation of fluidics consumables being replaced, the NovoExpress software will prompt a window to remind users to conduct the Priming procedure (Figure 1-10).

Figure 1-10 NovoExpress Software Message for Priming after Replacing Fluidic System Consumables

During the Priming procedure, the progress of the Priming will be shown in the NovoEx-press software (Figure 1-11).

Figure 1-11 NovoExpress Software Message for Priming in Process

The Priming procedure can also be manually initiated by clicking Instrument → Fluid-ics Maintenance → Priming on the NovoExpress software when the instrument is in Ready status.

Once the Priming procedure is completed, a dialog box will be prompted for conduct-ing the QC Test (Figure 1-12). Prepare the NovoCyte QC particle sample as described in Section 2.3.1 Prepare ACEA NovoCyte QC Particle Sample in NovoCyte® Flow Cytometer Operator’s Guide and place it in the tube holder. Click QC Test to initiate the QC Test. The QC Test will automatically start. Follow the steps and complete the QC Test. A QC Test must be conducted each time when fluidic system consumables are replaced in order to check the performance of the system.

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Figure 1-12 NovoExpress Software Message for Initiating the QC Test after Completion of Fluidic Sys-

tem Consumables Replacement

The following sections describe the detailed procedure for replacing fluidic system con-sumables. It is highly recommended to replace all the fluidic system consumables at the same time to ensure the optimal system performance. However, each component can also be replaced individually if necessary. Click Instrument → Replace Fluidic System Consum-ables in the NovoExpress software to initiate the process. Always conduct the Priming process after the fluidic system consumables have been replaced as prompted by the No-voExpress software or manually conduct the Priming process by clicking Instrument → Fluidics Maintenance → Priming.

► Replace the ACEA NovoFlow Sheath Fluid In-Line Filter

Open the front panel of the NovoCyte instrument, and locate the sheath fluid in-line filter shown in Figure 1-13.

Figure 1-13 Sheath Fluid In-line Filter

Procedure


2 Open the front panel of the instrument.

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3 Unscrew the Luer connectors at two ends of the filter to disconnect the tubings from the filter.

Use lint free wipes to gently wipe of the sheath fluid from the tubing and avoid sheath fluid dripping onto the instrument.

4 Replace the old filter with a new one and correctly connect the tubings to the filter.

Finger tightening the Luer connectors only to avoid damage of the filters and filter leakage.

5 Turn on the NovoCyte instrument, and automatic start-up cleaning process will be initi-ated. Observe the sheath fluid flows through the filter properly and make sure there is no leakage through the filter-tubing connections.

6 Once the instrument is Ready, Click Instrument → Fluidics Maintenance → Priming to prime the fluidic system.

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The ends of the sheath fluid in-line filter are designed to be different on purpose to ensure the in-line filter is connected into the system in the correct direction. Incorrect direction would lead to system malfunction.

When replacing the sheath fluid in-line filter, do not twist or kink the tubing.

► Replace the Waste Filter

Open the side panel of the NovoCyte instrument, and locate the waste filter as shown in Figure 1-14.

Figure 1-14 Waste Filter

Procedure


2 Open the side panel of the instrument.

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3 Unscrew the Luer connectors at two ends of the waste filter to disconnect the tubings from the filter.

Use lint free wipes to gently wipe of the sheath fluid from the tubing and avoid sheath fluid dripping onto the instrument.

4 Replace the old filter with a new one and correctly connect the tubing to the filter.

Finger tightening the Luer connectors only to avoid damage of the filters and filter leakage.

5 Turn on the NovoCyte instrument, and automatic start-up cleaning process will be initi-ated. Observe the waste flows through the waste filter properly and make sure there is no leakage through the filter-tubing connections.

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The ends of the waste filter are designed to be different on purpose to ensure the filter is connected into the system in the correct direction. Incorrect direction would lead to system malfunction.

When replacing the waste filter, do not twist or kink the tubing.

All biological samples or materials can contain infectious deadly disease. Please be careful and always wear protective clothing, masks, and gloves when handling the waste.

► Replace the ACEA NovoFlow Sheath Fluid Inlet Filter

ACEA NovoFlow sheath fluid inlet filter is placed inside the ACEA NovoFlow container to filter out micro-particles or impurities in the sheath fluid.

Procedure

1 Make sure the NovoCyte is in Ready status.

2 Press the metal clip on the quick coupler to disconnect the tubing from the ACEA Novo-Flow sheath fluid container.

3 Unscrew the cap and take the tubing-filter assembly out from the container and put it on a clean lint-free wipe (Figure 1-15).

Figure 1-15 Change ACEA NovoFlow Sheath Fluid Inlet Filter

4 Replace the old filter with a new one and correctly connect the tubing to the filter.

Please use proper force when tightening the Luer connectors.

The ends of the sheath fluid inlet filter are different to ensure the filter is connected into the system in the correct direction. Incorrect installation will lead to system malfunction.

5 Click Instrument → Fluidics Maintenance → Priming to prime the fluidic system.

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When replacing the sheath fluid inlet filter, do not twist or kink the tubing.

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► Replace the ACEA NovoRinse and ACEA NovoClean Solution Inlet Filter

ACEA NovoRinse and ACEA NovoClean solution inlet filters are placed inside the ACEA NovoRinse and ACEA NovoClean containers, respectively, to filter out micro-particles or impurities in the solution.

Procedure

1 Make sure the NovoCyte is in Ready status.

2 Press the metal clip on the quick coupler to disconnect the tubing from the ACEA Novo-Rinse or ACEA NovoClean container.

3 Unscrew the cap and take the tubing-filter assembly out from the container and put it on a clean lint free wipe. Take off the old filter and replace with a new one (Figure 1-16).

Figure 1-16 Change ACEA NovoRinse and ACEA NovoClean Solution Inlet Filter

4 Click Instrument → Fluidics Maintenance → Priming to prime the fluidic system.

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Unscheduled Maintenance

Remove Bubbles

2. Unscheduled Maintenance

The following instrument components should be checked occasionally and maintenance or cleaning should be conducted when necessary. The frequency depends on how often the cytometer is used. Other components not listed here should also be checked periodi-cally for wear and replacement if necessary.

Table 2-1 lists the unscheduled maintenance to be done on the NovoCyte instrument.

Unscheduled Maintenance When to Perform

Remove Bubbles ► The CV of the data is larger than normal

► QC test failed

Unclog the Sample Injection Probe (SIP)

► Fewer events are detected than expected

► Perform a large amount of tests in one day

Clean the External Surface of the Instrument

Keep the external surface of the instrument clean

Clean the Sample Injection Probe (SIP) The sample injection probe’s outer surface is dirty

Decontamination When suspecting bacteria contamination in the system

Clean Fluidic System before Long-Term Storage

No plan to run the NovoCyte cytometer for more than two weeks

Purge Fluidic System before Shipment The NovoCyte flow cytometer needs to be shipped

Calibrate the Fluidics Station Abnormal warning about the liquid level in the reagent containers is observed

Clean the Optical Filters and Mirrors The optical filters and mirrors are dirty

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Table 2-1 Unscheduled Maintenance and When to Perform

2.1 Remove Bubbles

When the CV of the data is larger than normal or the QC Test fails, there is a possibility that bubbles exist inside the flow chamber. In this case, the following procedure should be conducted to remove the bubbles.

When the NovoCyte instrument is in Ready status, place a tube containing at least 1 mL of 70% ethanol into the sample holder. Click Instrument → Fluidics Maintenance → De-bubble from the NovoExpress software to execute the debubble procedure.

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Unclog the Sample Injection Probe (SIP)

2.2 Unclog the Sample Injection Probe (SIP)

During a test, if much fewer events are detected than expected, there is possibility that the SIP is clogged. To solve this issue, when the NovoCyte instrument is in Ready status, click Instrument → Fluidics Maintenance → Backflush to execute the Backflush procedure to unclog the SIP.

If the problem still exists after Backflush, follow the procedure below to manually clear the clog inside the SIP.

Materials

► 1/16” Inner Diameter (ID) tubing (included in the NovoCyte maintenance kit)

► A 10 mL syringe


► ACEA NovoFlow sheath fluid

Procedure



3 Unscrew the connector to the SIP and remove the SIP from the instrument (Figure 2-1);

Please refer to Clean the Sample Injection Probe (SIP) of Section 1.1 Preventative Main-tenance.

Figure 2-1 Remove the SIP from the Instrument

Sample injection probe is fragile and may be damaged if handled improperly. When handling the sample injection probe, please be careful and prevent deforming or bending it.

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Clean the External Surface of the Instrument

4 Connect the SIP with the 1/16” ID tubing provided in the NovoCyte maintenance kit. Con-nect the other end of the tubing to a 10 mL syringe filled with ACEA NovoClean solution (1X) (Figure 2-2).

Figure 2-2 Connect the SIP with a 10 mL Syringe

5 Immerse the other end of the SIP into a beaker containing ACEA NovoClean solution (1X). Push the plunger of the syringe back and forth 10 times to flush the SIP with ACEA NovoClean solution.

6 Disconnect the SIP and merge it into a beaker containing ACEA NovoClean solution for about 10 minutes.

7 Immerse the SIP into a beaker containing ACEA NovoFlow sheath fluid for about 10 minutes.

8 Reinstall the SIP into the instrument.

9 Power on the instrument. Verify that the SIP is properly installed. If there is fluid leakage during the start-up process, please reinstall the SIP. If leakage persists, contact your local ACEA Biosciences representative for assistance.

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2.3 Clean the External Surface of the Instrument

Keep the external surface of the instrument clean using the following method.

Materials

► ACEA NovoRinse solution (1X)



Procedure

1 Turn off the power of the instrument and unplug the power cord.

To avoid the risk of electrical shock, make sure to power off the cytometer and unplug the power cord from the power outlet.

2 Gently wipe the surface of the instrument with a clean soft cloth soaked with ACEA NovoRinse solution (1X).

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Clean the Sample Injection Probe (SIP)

3 Wipe the surface again using a clean soft cloth.

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Biohazardous samples may make contact with the surface of the instrument. When cleaning the surface appropriate personal protective equipment should be worn.

Do not use isopropyl alcohol (IPA) or ethanol to clean the instrument surface.

2.4 Clean the Sample Injection Probe (SIP)

If the outer surface of the sample injection probe is dirty, follow the procedure below to manual clean the outer surface of the sample injection probe.

Materials




Procedure

1 Click Instrument → Shut Down on the NovoExpress software.

2 Select Clean sample injection probe option and then click Yes button in the prompted window (Figure 2-3). NovoExpress software will guide you to conduct the cleaning SIP procedure.

Figure 2-3 Message in NovoExpress Software when Shutting down

The instrument will shut down directly if Clean sample injection probe option is not selected.

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Clean the Sample Injection Probe (SIP)

3 Follow the instruction in the software, remove the tube from the tube holder and click OK button (Figure 2-4).

Figure 2-4 Message in NovoExpress Software to Conduct the SIP Cleaning Procedure

Sample injection probe will move downward after clicking the OK button. Do not put your fingers underneath the sample injection probe!

4 After clicking the OK button, the sample injection probe will move downwards. Prepare a damp soft cloth with ACEA NovoClean solution (1X) and gently wipe the outer surface of the sample injection probe.


Sample injection probe is fragile and may be damaged if handled improperly. When cleaning the sample injection probe, please be careful and prevent deforming or bending it.

5 Click OK button after cleaning. (Figure 2-5).


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Decontamination

6 Please put a tube of filtered DI water or ACEA NovoRinse solution (1X) in the sample holder and click OK button again ( Figure 2-6). The instrument will shut down automati-cally.


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2.5 Decontamination

The decontamination procedure can be used either after a known bacterial contamination has occurred, or as a preventative maintenance procedure to minimize or prevent the oc-currence of bacterial contamination in the NovoCyte fluidics system.

A Version 2 Decontamination Kit (Catalog No. 2030016) is required to run this procedure.

The Decontamination function is only available by logging in as account with the De-contaminate Instrument privilege.

Procedure

1 Click Instrument → Fluidics Maintenance → Decontamination from the NovoExpress soft-ware.

2 NovoExpress software will prompt a dialog box to guide you conducting the decontami-nating procedure (Figure 2-7). Follow the instructions in each step and click OK to move to the next step until completion.

3 The NovoCyte flow cytometer is ready to use.

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Clean Fluidic System before Long-Term Storage

Figure 2-7 NovoExpress Dialog Box to Guide the Decontamination Procedure

Contact your local ACEA Biosciences representative or distributor for any questions regarding this procedure.

2.6 Clean Fluidic System before Long-Term Storage

If there is no plan to run the NovoCyte flow cytometer for more than two weeks, a thor-ough cleaning process is recommended to prevent salt crystallization in the fluidic system.

Procedure

1 When the cytometer is in Ready status, replace all the instrument reagents with deion-ized water.

2 Click Instrument → Fluidics Maintenance → Priming from the NovoExpress software. Re-peat the procedure two times.

3 Press the power switch on the front panel of the NovoCyte instrument to initiate the au-tomatic shutdown cleaning process.

4 After the shutdown cleaning process is completed, the instrument will power off au-tomatically. The NovoExpress software status bar will show Instrument not Connected message.

5 Empty all the reagent containers and empty the waste. Rinse the containers with DI water thoroughly and air dry.

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When using the system after long-term storage, follow the procedure below to prepare the system.

1 Sterilize the reagent containers following the procedure described in Sterilize Instrument Reagent Containers of Section 1.1 Preventative Maintenance.

2 Add the ACEA NovoFlow, ACEA NovoRinse, and ACEA NovoClean to the corresponding containers.

3 Press the power button on the front panel of the NovoCyte instrument. The instrument will start the automatic start-up cleaning procedure.

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Purge Fluidic System before Shipment

4 Click Instrument → Fluidics Maintenance → Priming from the NovoExpress software. Re-peat the procedure two times.

5 The NovoCyte instrument is then ready to use.

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If NovoCyte instrument has not been used for more than two months, it is recommend-ed to replace the fluidic system consumables before use. Refer to Section 1.2 Replacing Fluidic System Consumables in this guide for detailed procedure.

2.7 Purge Fluidic System before Shipment

If for any reason the NovoCyte flow cytometer needs to be shipped, perform the following procedure to purge the fluidic system before packaging and shipping

The Purge function is only available by logging in as account with the Purge Instrument privilege.

Procedure

1 Click Instrument → Fluidics Maintenance → Purge from the NovoExpress software.

2 NovoExpress software will prompt a dialog box to guide you to conduct the purging procedure (Figure 2-8). Follow the instructions in each step and click OK to move to the next step until completion.

Figure 2-8 NovoExpress Dialog Box to Guide the Purge Procedure

3 Power off the instrument. Unplug the power supply and accessories.

4 Sterilize the reagent containers following the procedure described in Sterilize Instrument Reagent Containers of Section 1.1 Preventative Maintenance.

5 Pack the instrument and accessories properly for shipment.

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If there are operating questions when purging the fluidics system, in order to avoid dam aging the instrument, please stop purging and contact your local ACEA Biosciences representative or distributor.

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Calibrate the Fluidics Station

2.8 Calibrate the Fluidics Station

Fluidics station is calibrated at the factory and should be ready to use when connected to NovoCyte instrument after installation. However, in some cases there is a need to recali-brate the fluidics station after shipment and movement. When a false warning about the liquid level in the reagent containers is observed, follow the procedures below to recali-brate the fluidics station.

Calibrate Fluidics Station function is only available when login as account with the Calibrate Fluidics Station privilege.

Procedure

1 Click Instrument → Operation → Calibrate Fluidics Station in the NovoExpress software.

Figure 2-9 NovoExpress Software Instrument Menu

2 Follow the instruction in the software, remove the ACEA NovoFlow, ACEA NovoRinse, ACEA NovoClean, and waste containers from the fluidics station and click the OK button (Figure 2-10);

Figure 2-10 Message in NovoExpress Software when Performing Fluidics Station Calibration

3 The software will prompt a dialog box indicating the fluidics station calibration is suc-cessful. Follow the instruction to place the ACEA NovoFlow, ACEA NovoRinse, ACEA NovoClean, and waste containers back into the fluidics station and click the OK button (Figure 2-11);

Figure 2-11 NovoExpress Software Message Indicating Fluidics Station Calibration is Successful

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Clean the Optical Filters and Mirrors

2.9 Clean the Optical Filters and Mirrors

In normal operation, the optical filters and mirrors are tightly enclosed inside the optical fixtures and should remain clean. However, it might get dirty if unplugged from the slot or be replaced. The following method describes how to clean the optical filters or dichroic mirrors when it gets dirty. Maintaining the optical filters and mirrors surface clean is very important to get the good quality signals.

When cleaning or replacing optical filters or dichroic mirrors, handle with care to avoid scratching the surface and to prevent the filters from falling out of the holder. Use cotton swabs, optical lens paper, and spectral-grad methanol or analytical grade ethanol in a dropper bottle to clean the optical filters. Acetone should not be used for cleaning the optical filters. Always wear laboratory gloves when handling the optical filter.

Materials

► Analytical grade ethanol

► Compressed air or a bulb blower

► Lint-free cotton swabs

► Lint-free wipe or lens cleaning tissue

Procedure

1 Blow off the contaminants

Many contaminants are just loosely attached to the surface of the optical filters and can be blown off. Wear clean laboratory gloves and hold the filter firmly in one hand. Always use a gentle to moderate air flow and maintain an oblique angle. Slowly move the air steam over the filter surface at an angle to gently blow off the contaminants. Repeat this process for the other side of the filter.

2 Wrap a triangular-shaped lint-free wipe or lens cleaning tissue around a lint-free cotton

swab. Moisten and seal the end with a few drops of alcohol.

3 Holding the cotton swab in a horizontal position, gently clean the surface of the filter with a continuous motion at a constant speed.

4 Allow the solvent to evaporate. Use a room light to inspect the filter to ensure it is clean. If contamination remains, start with a brand new wipe and swab and repeat step 2-3 again.

5 Repeat step 1-4 for the other side of the filter.

6 Insert the cleaned filter into the filter slot according to the correct orientation and push it all the way in.

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Always blow the contaminants off the filters before using the wipe or swab to clean the filter surface. Failure to do so may scratch the filter surface.

Do not reuse the wipes and swabs. Always use new wipes and swabs for every new cleaning attempt.

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Troubleshooting

3. Troubleshooting

This section contains troubleshooting guidelines. Follow the procedures described in this section for troubleshooting methods.

For technical support, please refer to the Prologue IX: Technical Support section in this guide.

Refer to the NovoExpress® Software Guide for related issues with the software.

Refer to the NovoSampler® Operator’s Guide for NovoSampler troubleshooting.

Refer to the NovoSampler® Pro Operator’s Guide for NovoSampler Pro troubleshooting.

Table 3-1 provides guidance on troubleshooting the NovoCyte instrument, NovoSampler and NovoSampler Pro. Table 3-2 lists the message IDs and the associated software prompted mes-sages.

Please contact ACEA Technical Support if issues are not resolved after performing the recom-mended solutions or if there are any issues which are not listed in these two tables.

Investigate possible causes and perform the recommended solutions in the given order.

Observation Possible Causes Recommended Solutions

Instrument QC test fails QC particles are expired or not stored appropri-ately

Prepare a new tube of QC particles and mix them well to run the QC test again.

Flow cell is dirty Perform the maintenance functions from the NovoExpress software: Unclog, Priming, Extensive Rinse, and Extensive Clean.

Air bubbles exist in the flow chamber

Execute Debubble and Priming function from NovoExpress Software.

Zero or low number of events detected

Threshold is set too high Lower the threshold setting and re-collect the sample.

Sample concentration is too low

Redo sample preparation using an appro-priate method to concentrate the sample.

Sample is not mixed well Mix the sample well to suspend the cells.

NovoFlow container is empty

Refill the NovoFlow container following the procedures described in Section 2.1.2 Add Instrument Reagents in NovoCyte® Flow Cy-tometer Operator’s Guide.

NovoFlow container is not correctly connected to the instrument and the NovoFlow solution does not flow through the tubing

► Reconnect the NovoFlow quick coupling connectors to ensure the NovoFlow container is correctly con-nected to the instrument.

► Execute Prime function from the Novo-Express Software.

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Troubleshooting


Zero or low number of events detected

Sample Injection Probe is clogged

► Execute the Backflush function from the NovoExpress Software.

► Follow the procedures described in Section 2.2 Unclog the Sample Injection Probe (SIP) in this guide to unclog the sample injection probe.

Flow cell is clogged Execute the Unclog function from the No-voExpress Software.

Sample tubing is leaking Clean the leaked fluid with wipes and properly reconnect the sample tubing.

Air bubbles exist in the Sampling Pump

Execute Debubble and Priming function from NovoExpress Software.

High CV Air bubbles exist in the flow chamber

Execute the Debubble and Priming function from NovoExpress Software.

Sampling tubing is clogged

Perform maintenance functions from No-voExpress software: Unclog, Priming, Ex-tensive clean, Extensive rinse.

Sheath in-line filter is clogged

Replace the sheath in-line filter following the procedures described in Section 1.2 Replacing Fluidic System Consumables in this guide.

Improper sample prepa-ration

Re-prepare the sample properly.

The sample flow rate is set too high

Lower the sample flow rate setting in the NovoExpress software.

Excessive debris in dis-play

Sheath in-line filter is dirty

Replace the sheath in-line filter following the procedures described in Section 1.2 Replacing Fluidic System Consumables in this guide.

The threshold is set too low

Increase the threshold setting and re-col-lect the sample.

Fluidic system tubing is dirty

► Execute Rinse function from NovoEx-press Software.

► Execute Decontamination function from NovoExpress Software.

High event rate The threshold is set too low

Increase the threshold setting and re-col-lect the sample.

Sample concentration is too high

Re-prepare the sample to appropriate con-centration, or dilute the sample.

Sample flow rate is too high

Reduce the sample flow rate.

NovoExpress displays In-strument not connected even though the instru-ment is powered up

Workstation is in hi-bernation causing the interruption of commu-nication between the NovoCyte instrument and workstation

Restart NovoExpress Software.

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Troubleshooting


NovoExpress displays In-strument not connected even though the instru-ment is powered up

Improper connection between the NovoCyte instrument and worksta-tion

Reconnect the USB cable between the in-strument and the workstation.

More than one NovoEx-press instances are run-ning.

Only run one instance of NovoExpress soft-ware.

Unstable Signal or gap in the data plot

Air bubbles exist in the flow chamber

Execute the Debubble and Priming function from NovoExpress Software.

Sample tube does not fit the sample tube holder

Wrong type of sample tube is used

Use 12 × 75 mm flow tube or 1.5 mL Ep-pendorf tube.

Sample Injection Probe is not properly aligned with the wells or tubes

Plate or tube rack is not loaded correctly.

Load the plate or tube rack with the cor-rect orientation. Make sure the plate or tube rack is seated onto the sample tray properly.

NovoSampler (Pro) is not calibrated.

Click Instrument → NovoSampler (Pro) → Calibrate in the NovoExpress Software to re-calibrate the NovoSampler (Pro).

Leakage from the SIP Waste tubing is de-formed or kinked

► Reconnect the waste tubing to ensure it is properly connected.

► Move the instrument forward as needed to ensure sufficient space bet-ween the instrument and wall. Refer to Section 1.2.1 Installation in NovoCyte® Flow Cytometer Operator’s Guide for more information on properly installing NovoCyte instrument.

Waste filter is clogged Replace the waste filter following the pro-cedures described in Section 1.2 Replacing Fluidic System Consumables in this guide.

Leakage from Sampling Pump

Incorrect tubing con-nection to the sampling pump

Reconnect the tubing to the sampling pump.

Leakage from other components of the in-strument

Loose fluidic compo-nents connection or or blocked sheath in-line filter

► Power off the NovoCyte instrument.

► Clean the leaked fluid.

► Replace the sheath in-line filter following the procedures described in Section 1.2 Replacing Fluidic System Consumables in this guide.

► Contact ACEA technical support for service.

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Table 3-1 Troubleshooting Guide for NovoCyte Instrument

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Troubleshooting

Message IDs

Software Messages Possible Causes Recommended Solutions

0x0001 Collision of SIP The movement of SIP is blocked by some obstacles

Locate and clear the obsta-cles. Click the OK button in the prompted dialog box or wait 10 seconds for automatic error han-dling. The instrument will move the SIP to the home position. This procedure will take about 3 min-utes.

Incorrect plate is se-lected in plate man-ager

Select the correct plate in the plate manager window.

Incorrect positioning of plate in NovoSam-pler (Pro)

► Position the plate on the shaker correctly. Ensure the plate is seated flat on the stage inside the clamps.

► Re-calibrate NovoSampler Pro.

Dirty SIP or SIP wash apparatus

Clean the SIP or SIP wash ap-paratus following the procedures described in Section 1.1 Preven-tative Maintenance in this guide.

0x0002 Running out of NovoFlow NovoFlow solution is not sufficient to continue to run any samples

Refill the NovoFlow container fol-lowing the procedures described in Section 2.1.2 Add Instrument Reagents in NovoCyte® Flow Cy-tometer Operator’s Guide.

Fluidic station is not working properly

Replace the fluidic station.

0x0003 Running out of Novo-Rinse

NovoRinse solution is not sufficient to continue to run any samples

Refill the NovoRinse container following the procedures de-scribed in Section 2.1.2 Add In-strument Reagents in NovoCyte® Flow Cytometer Operator’s Guide.



0x0004 Running out of Novo-Clean

NovoClean solution is not sufficient to continue to run any samples

Refill the NovoClean container following the procedures de-scribed in Section 2.1.2 Add In-strument Reagents in NovoCyte® Flow Cytometer Operator’s Guide.



0x0005 Waste container is full Waste container is too full to continue to run any samples

Empty the waste container fol-lowing the procedures described in Section 2.1.3 Empty Waste in NovoCyte® Flow Cytometer Op-erator’s Guide.



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Troubleshooting

Message IDs


0x0006 System voltage is out of range

System Error Restart the NovoCyte instrument.

0x0007 System electric current is out of range


0x0008 Firmware configuration error


0x0009 0x000A0x000B

…… laser self test error Specified laser is not functioning properly

The instrument will automatically reset the laser and run a laser self-test. It takes approximately 5 to 10 minutes.

0x000C 0x000D0x000E

…… laser is not con-nected

Specified laser is not detected

Restart the NovoCyte instrument.

0x000F NovoSampler communi-cation lost

The cable between the NovoSampler (Pro) and NovoCyte flow cytometer is not securely connected

Reconnect the cable between the NovoSampler (Pro) and Novo-Cyte flow cytometer.

NovoSampler (Pro) is not communicat-ing with NovoCyte

Restart NovoCyte instrument.

0x0010 NovoSampler (Pro) has not been calibrated

NovoSampler (Pro) is newly installed or re-connected.

Follow the prompted instructions to calibrate the NovoSampler (Pro).

0x0011 NovoSampler (Pro) cali-bration failed……

NovoSampler (Pro) is not installed prop-erly

Re-install and calibrate the No-voSampler (Pro) following the procedures described in Section 1.2 Installation in NovoSampler® Operator’s Guide or NovoSam-pler® Pro Operator’s Guide.

The NovoSampler (Pro) cover was opened during cali-bration.

Close the NovoSampler (Pro) cover and redo the calibration.

0x0012 The movement of plate is out of range

The movement of the orbital shaker is blocked.

► Check the path of the orbital shaker to make sure there are no objects blocking the movement. Clear the block if there is any.

► Click Instrument → No-voSampler Pro → Calibrate in NovoExpress Software to re-calibrate the NovoSam-pler (Pro).

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Troubleshooting

Message IDs


0x0012 The movement of plate is out of range

NovoSampler (Pro) is not installed prop-erly

Re-install and calibrate the No-voSampler (Pro) following the procedures described in Section 1.2 Installation in NovoSampler® Operator’s Guide or NovoSam-pler® Pro Operator’s Guide.

0x0013 Cover of NovoSampler (Pro) is opened during moving plate

The cover of No-voSampler (Pro) is opened during mov-ing plate

Close the NovoSampler (Pro) cover. NovoSampler (Pro) will au-tomatically reset and be ready for operation.

0x0014 Pressure is out of limit Waste container is not correctly con-nected to the instru-ment

Check the quick coupling con-nectors to ensure the waste con-tainer is correctly connected to the instrument.


Replace the sheath in-line filter following the procedures de-scribed in Section 1.2 Replacing Fluidic System Consumables in this guide.

Flow cell is clogged Perform maintenance procedures from NovoExpress software, Un-clog, Priming, Extensive Clean and Extensive rinse.

0x0015 NovoSampler firmware error

NovoSampler (Pro) firmware is not working properly

Re-install or upgrade the No-voSampler (Pro) firmware.

0x0016 0x0017 0x0018

…… laser does not emit Specified laser is not connected properly or laser is not work-ing properly

► Reconnect the laser properly.

► Restart the instrument.

0x0019 0x001A 0x001B

…… laser communica-tion error

Interference or bad connection or bad laser

► Reconnect the laser properly.

► Restart the instrument.

0x001C Sample injection probe reset failed

Bad connection or optocoupler is dam-aged

Click the OK button in the prompted dialog box or wait 10 seconds for automatically error handling. Restart the instrument.

0x001D Sampling Pump reset failed


Restart the instrument.

0x001E The SIP module needs to be upgraded to support NovoSampler Pro

SIP module is not compatible with con-nected NovoSampler Pro

Contact ACEA technical support for service.

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Troubleshooting

Message IDs


0x0020 System initialization is paused

Liquid level in the reagent containers is not within normal range when Novo-Cyte is powered up.

► Make sure that the instru-ment reagent containers are placed correctly and the liquid level in the containers is within the normal range.

► Click OK on the prompted dialog to continue system initialization.

0x0021 Sheath fluid pump reset failed


Restart the instrument.

0x0022 Resetting NovoSampler Pro to zero position failed

NovoSampler Pro is not working prop-erly.

Click Reset. Restart the No-voSampler Pro.

0x0023 Resetting NovoSampler Pro to home position failed

NovoSampler Pro is not working prop-erly.

Click Reset. Restart the No-voSampler Pro.

0x0100 Instrument cover opened Instrument cover is opened or not tightly closed

Close the instrument cover.

0x0101 NovoFlow running low NovoFlow solution is below the pre-set volume limit

Refill the NovoFlow container fol-lowing the procedures described in Section 2.1.2 Add Instrument Reagents in NovoCyte® Flow Cy-tometer Operator’s Guide.

Fluidic station not working properly


0x0102 NovoRinse running low NovoRinse solution is below the pre-set volume limit

Refill the NovoRinse container following the procedures de-scribed in Section 2.1.2 Add In-strument Reagents in NovoCyte® Flow Cytometer Operator’s Guide.


Replace fluidic station.

0x0103 NovoClean running low NovoClean solution is below the pre-set volume limit

Refill the NovoClean container following the procedures de-scribed in Section 2.1.2 Add In-strument Reagents in NovoCyte® Flow Cytometer Operator’s Guide.



0x0104 Waste container is close to full

Waste is above the pre-set volume limit

Empty the waste container fol-lowing the procedures described in Section 2.1.3 Empty Waste in NovoCyte® Flow Cytometer Op-erator’s Guide.



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Troubleshooting

Message IDs


0x0105 Cover of NovoSampler (Pro) is opened

Cover of NovoSam-pler (Pro) is opened

Close the cover.

0x0106 NovoSampler (Pro) is disconnected when powered up

The NovoSampler (Pro) is disconnect-ed when powered up

► Shut down NovoCyte Flow Cytometer.

► Reconnect the cable of the NovoSampler (Pro) to Novo-Cyte flow cytometer.

► Turn on NovoCyte and follow the prompts to calibrate the NovoSampler (Pro).

0x0109 Fluidics station is not connected

Cable between the fluidics station and the NovoCyte instru-ment is not properly connected

► Power down the instrument.

► Reconnect the cable bet-ween the fluidics station and the NovoCyte instrument.

► Power up the instrument.

0x010A 0x010B 0x010C 0x010D

…… liquid level sensor failure

Specified liquid level sensor in the fluidics station is not work-ing properly

► Reconnect the fluidics stati-on cable.

► Restart the NovoCyte instru-ment.

► Replace the fluidics station.

0x010F Sheath filter is clogged. Please replace the sheath filter and run the Priming procedure


Replace the sheath in-line filter following the procedures de-scribed in Section 1.2 Replacing Fluidic System Consumables in this guide.

0x0110 Recovering collision error Plate stops at an in-correct position

No action is needed. Instrument will automatically recover the er-ror.

0x0111 Orbital shaker homing position reset failure

Orbital shaker of No-voSampler Pro is not working properly

No action is needed. The No-voSampler Pro can be used nor-mally.

Communication error (code: xx, xx). Please restart NovoCyte and NovoExpress

USB cable between the NovoCyte instru-ment and worksta-tion is not connected properly

► Reconnect the USB cable between the NovoCyte instrument and the worksta-tion.

► Restart the instrument and NovoExpress software.

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Table 3-2 NovoExpress Prompted Error Messages and Message IDs

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