NovoCyte Flow Cytometer. Maintenance Guide

Transcription

1 NovoCyte Flow Cytometer Maintenance Guide V1

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3 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 Scheduled Maintenance Preventative Maintenance Replacing Fluidic System Consumables Unscheduled Maintenance Remove Bubbles Unclog the Sample Injection Probe (SIP) Clean the External Surface of the Instrument Clean the Sample Injection Probe (SIP) Decontamination Clean Fluidic System before Long-Term Storage Purge Fluidic System before Shipment Calibrate the Fluidics Station Clean the Optical Filters and Mirrors Troubleshooting 32 3

4 4 Table of Content

5 Prologue Revision History Prologue I. Revision History Version Revision Date Copyright 2017, 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 related 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 software 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 latest 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 improvement. 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 Mesa Ridge Rd. #100 San Diego, CA USA 5

6 Prologue Contact Information II. Contact Information Manufacturer China Distributor International Distributors ACEA Biosciences, Inc Mesa Ridge Rd., Suite 100 San Diego, CA USA ACEA Bio (Hangzhou) Co., Ltd. Building 5, 2# Xiyuan Wu Rd., Xihu District Hangzhou, Zhejiang Province, P. R. China Please contact ACEA Biosciences, Inc., or visit for a complete list of international distributors III. Trademark ACEA 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 Guide This maintenance guide describes the maintenance of the ACEA NovoCyte flow cytometer. Most of these functions are controlled by the ACEA NovoExpress software. For additional information about the ACEA NovoCyte, please refer to NovoCyte Flow Cytometer 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 Numbered Listing Description Describes the steps in a procedure that must be performed in the listed order. 6

7 Prologue Technical Support Format Italic font, gold Italic font Ctrl+X Description Points to a different chapter in this guide, which should be referred to for better understanding. Describes buttons, icons or functions when operating the NovoExpress software. In addition, important notes and information are also shown in italic font. Indicates the sequence of the menu operation in NovoExpress software. For example, File Print means to select the Print function from the File menu. 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 Laser Radiation Warning Shock Hazard This symbol indicates that certain precautions must be taken when working with potentially infectious biological specimens and materials. This symbol indicates that the operation could potentially cause exposure to laser radiation. Protective measures should be taken during the operation. This symbol indicates that improper or unsafe operation could result in instrument damage, data loss, and personal injury. This symbol indicates that the operation has a risk of electric shock. Important Note Additional Information This symbol indicates information which is critical to the success of the procedure or use of the product. This symbol provides additional information about the current topic or procedure. uuu Form Table continues on next page. The End of the Table End of a table. VI. Technical Support If there are operating or technical questions, please refer to the sections relating to the instrument operation in this guide. For additional technical support, please contact your local 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. 7

8 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 For support within China, please call For users in other countries or regions, contact your local ACEA Biosciences representatives or distributors, which may be found at When encountering an issue during operation of NovoCyte flow cytometer, NovoSampler 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 experiment file and other information that helps in the diagnosis and troubleshooting of NovoCyte instrument. User can also attach any other files using this function. Please refer to the NovoExpress Software Guide for detailed information on this function. 8

9 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 incorporates automatic procedures for maintenances as much as possible, eliminating timeconsuming manual maintenance. This chapter describes the procedures to execute or conduct the maintenance procedures. Table 1-1 lists the regular maintenance to be done on the NovoCyte instrument. Maintenance Daily Start-up Daily Shutdown Recommended Maintenance Frequency Daily. Automatic system cleaning during start-up. Daily. Automatic system cleaning and decontamination during shutdown. 1 Add Instrument Reagents (ACEA NovoFlow Sheath Fluid, ACEA NovoRinse Solution, and ACEA NovoClean Solution) Empty Waste Preventative Maintenance Clean the Sample Injection Probe (SIP) Clean the SIP Cleaning Apparatus Conduct Extensive Rinse Unclog the Flow Cell Sterilize Instrument Reagent Containers Replacing Fluidic System Consumables Replace the ACEA Sheath Fluid In-Line Filters Replace the Waste Filter Replace the Sheath Pump Tubing Replace the ACEA NovoFlow Sheath Fluid Inlet Filter Replace the ACEA NovoRinse and ACEA NovoClean Solution Inlet Filters As needed. Check the instrument reagent volume in the containers before each experiment and add more reagents when necessary. As needed. Check waste volume before each experiment and empty waste container when necessary. Every month Every two months or Prompted by NovoExpress Software Table 1-1 Scheduled Maintenance and Recommended Frequency 9

10 Scheduled Maintenance Preventative Maintenance 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) 1 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 uuu 10

11 Scheduled Maintenance Preventative Maintenance 4 Unscrew the white connector by turning it counter clockwise. 1 Figure 1-2 Unscrew the White 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 white connector from the slot. Handle the assembly 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 uuu 11

12 Scheduled Maintenance Preventative Maintenance 5 Remove the SIP straight up from the housing slot. 1 Figure 1-4 Take SIP out from the Housing Slot Sample injection probe directly contacts the biological samples which may be biohazardous. Wear gloves when handling 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. 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 white connector and tubing by finger tightening it clockwise. uuu 12

13 Scheduled Maintenance Preventative Maintenance 10 Power on NovoCyte instrument and start NovoExpress software. After the instrument is Ready, click Instrument Fluidics Maintenance Backflush to flush the sample injection probe. Clean the SIP Cleaning Apparatus Materials Clean soft cloth ACEA NovoClean solution (1X) Deionized (DI) water Procedure 1 1 Turn off the power of the instrument. 2 Open the top cover of the instrument. 3 Prepare a damp cloth with ACEA NovoClean solution (1X). Wipe the SIP cleaning apparatus 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 injection probe. SIP cleaning apparatus directly contacts the biological samples which may be biohazardous. 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 in Ready status, click the Instrument Fluidics Maintenance Extensive Rinse from NovoExpress software, and the Extensive Rinse procedure will automatically be executed. 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 Unclog from NovoExpress software, and the Unclog procedure will automatically be executed. 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. 13

14 Scheduled Maintenance Replacing Fluidic System Consumables Materials Clean soft cloth ACEA NovoClean solution (1X) Deionized (DI) water (or ACEA NovoFlow solution) Procedure 1 Wipe the outer surface of the container with a clean soft cloth. 1 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. 1.2 Replacing Fluidic System Consumables The NovoCyte flow cytometer monitors the accumulated running time of the fluidic system 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 1-6). Click Replace to continue. Clicking the Close button will cancel the process and allow continued use of the system. However the consumables should be replaced promptly. Figure 1-6 NovoExpress Software Message for Replacing Fluidic System Consumables Click the Replace button and a dialog box as shown in Figure 1-7 will be prompted. The replacement of the sheath pump tubing requires the input of the Tubing Calibration Number, which is the last three digits of the tubing serial number affixed to the tubing (Figure 1-7). Enter the Tubing Calibration Number in the prompted dialog box and click Next to continue. 14

15 Scheduled Maintenance Replacing Fluidic System Consumables Figure 1-7 NovoExpress Software Dialog Box for Entering the Tubing Calibration Number 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. 1 Make sure the Tubing Calibration Number is input correctly. An incorrect Tubing Calibration Number will result in incorrect sheath flow rate and affects the performance of the system. 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 If no Tubing Calibration Number is found on the sheath pump tubing, click Next to continue and a dialog box as shown in Figure 1-9 will be prompted. The system will then use a default value for the Tubing Calibration Number. Clicking Next will continue the process using the default value. Figure 1-9 NovoExpress Software Message when No Tubing Calibration Number is Entered It is strongly recommended to input the Tubing Calibration Number labelled on the tubing. Click the Back button in Figure 1-9 to enter the correct Tube Calibration Number. Only if the Tubing Calibration Number is not immediately available, one may go ahead to replace the pump tubing without entering the Tubing Calibration Number. 15

16 Scheduled Maintenance Replacing Fluidic System Consumables NovoExpress software will then prompt the message shown in Figure Figure 1-10 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 (2pcs) (Cat. No ) Waste filter (1pcs) (Cat. No ) Sheath pump tubing (1pcs) (Cat. No ) ACEA NovoFlow sheath fluid inlet filter (1pcs) (Cat. No ) ACEA NovoRinse solution inlet filter (1pcs) (Cat. No ) ACEA NovoClean solution inlet filter (1pcs) (Cat. No ) 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. A confirmation message will be prompted (Figure 1-11). Click Yes to inform the system that the consumables have been replaced. Figure 1-11 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 1-12). Figure 1-12 NovoExpress Software Message for Priming after Replacing Fluidic System Consumables 16

17 Scheduled Maintenance Replacing Fluidic System Consumables During the Priming procedure, the progress of the Priming will be shown in the NovoExpress software (Figure 1-13). Figure 1-13 NovoExpress Software Message for Priming in Process The Priming procedure can also be manually initiated by clicking Instrument Fluidics Maintenance Priming on the NovoExpress software when the instrument is in Ready status. 1 Once the Priming procedure is completed, a dialog box will be prompted for conducting the QC Test (Figure 1-14). Prepare the NovoCyte QC particle sample as described in Section Prepare ACEA NovoCyte QC Particle Sample of 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. Figure 1-14 NovoExpress Software Message for Initiating the QC Test after Completion of Fluidic System Consumables Replacement 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 the optimal system performance. However, each component can also be replaced individually if necessary. Click Instrument Replace Fluidic System Consumables 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 NovoExpress software or manually conduct the Priming process by clicking Instrument Fluidics Maintenance Priming. 17

18 Scheduled Maintenance Replacing Fluidic System Consumables Replace the ACEA NovoFlow Sheath Fluid In-Line Filter Open the front panel of the NovoCyte instrument, and locate the two sheath fluid in-line filters shown in Figure Figure 1-15 Sheath Fluid In-line Filters Procedure 1 Make sure the NovoCyte is in Ready status. 2 Open the front panel of the instrument. 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 off the sheath fluid from the tubing and avoid sheath fluid dripping onto the instrument. 4 Replace the old filters with new ones and correctly connect the tubing to the filters. Finger tightening the Luer connectors only to avoid damage of the filters and filter leakage. 5 Click Instrument Fluidics Maintenance Priming to prime the fluidic system. Check that the sheath fluid flows through the two filters properly and make sure there is no leakage through the filter-tubing connections. The ends of the sheath fluid in-line filter are different to ensure the in-line filter is connected into the system in the correct direction. Incorrect installation will lead to system malfunction. When replacing the sheath fluid in-line filter, do not twist or kink the tubing. 18

19 Scheduled Maintenance Replacing Fluidic System Consumables Replace the Waste Filter Open the side panel of the NovoCyte instrument and locate the waste filter as shown in Figure Figure 1-16 Waste Filter Procedure 1 Make sure the NovoCyte is in Ready status. 2 Open the side panel of the instrument. 3 Unscrew the Luer connectors at two ends of the waste filter to disconnect the tubing 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 filters. Finger tightening the Luer connectors only to avoid damage of the filters and filter leakage. 5 Click Instrument Fluidics Maintenance Priming to prime the fluidic system. Check that the waste flows through the waste filter properly and make sure there is no leakage through the filter-tubing connections. Please install the waste filter in the correct direction as indicted by the inlet label on the filter and the flow direction arrows labeled on the tubing. Incorrect installation will lead to system malfunction. When replacing the waste filter, do not twist or kink the tubing. Biological samples may be biohazardous. Appropriate personal protective equipment should be worn when handling waste, in accordance with local safety regulations. Replace the Sheath Pump Tubing The sheath pump tubing should only be replaced using the Replace Fluidics Consumables function described above. The replacement of the sheath pump tubing requires the input of the Tubing Calibration Number that is affixed to sheath pump tubing. Failure to input the Tubing Calibration Number will affect the performance of the system. 19

20 Scheduled Maintenance Replacing Fluidic System Consumables Open the side panel of the NovoCyte instrument to access the sheath pump as shown in Figure Figure 1-17 Sheath Pump Procedure 1 Make sure the NovoCyte is in Ready status. 2 Open the side panel of the instrument. 3 Unscrew the Luer connectors at two ends of the sheath pump tubing. 4 While grabbing two sides of the pump retainer clip, gently squeeze in and remove the clip from the pump head (Figure 1-18). Figure 1-18 Remove the Sheath Pump Retainer Clip uuu 20

21 Scheduled Maintenance Replacing Fluidic System Consumables 5 Pull the Luer connectors outward and slide the luer lock fittings off the pump head (Figure 1-19). 1 Figure 1-19 Slide Luer Lock Fittings off the Pump Head 6 Remove the sheath pump tubing from the pump head. 7 Replace with a new sheath pump tubing by sliding the Luer lock fittings onto the pump head (Figure 1-20). Figure 1-20 Place a New Sheath Pump Tubing onto the Pump Head 8 Snap the pump retainer clip into pump head. Reconnect the Luer connectors of the pump tubing. The arrows labeled on the pump head and the tubing indicates the direction of fluid flow. Connect the tubing to the pump head in the correct direction as indicated. Incorrect direction will result in instrument malfunction. Finger tightening the Luer connectors only to avoid damage of the connectors. Ensure the sheath pump tubing and the fluidic tubing are not twisted or kinked. 9 Click Instrument Fluidics Maintenance Priming to prime the fluidic system. Check that the sheath fluid flows through the sheath pump properly and make sure there is no leakage through the pump tubing. 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. 21

22 Scheduled Maintenance Replacing Fluidic System Consumables 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 1-21). 1 Figure 1-21 Change ACEA NovoFlow Sheath Fluid Inlet Filter 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 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. When replacing the sheath fluid inlet filter, do not twist or kink the tubing. 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 1-22). Figure 1-22 Change ACEA NovoRinse and ACEA NovoClean Solution Inlet Filter 4 Click Instrument Fluidics Maintenance Priming to prime the fluidic system. 22

23 Unscheduled Maintenance Remove Bubbles 2. Unscheduled Maintenance 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. Table 2-1 lists the unscheduled maintenance to be done on the NovoCyte instrument. Unscheduled Maintenance Remove Bubbles Unclog the Sample Injection Probe (SIP) Clean the External Surface of the Instrument Clean the Sample Injection Probe (SIP) Decontamination Clean Fluidic System before Long-Term Storage Purge Fluidic System before Shipment Calibrate the Fluidics Station Clean the Optical Filters and Mirrors When to Perform The CV of the data is larger than normal QC test failed Fewer events are detected than expected Perform a large amount of tests in one day Keep the external surface of the instrument clean The sample injection probe s outer surface is dirty When suspecting bacteria contamination in the system No plan to run the NovoCyte cytometer for more than two weeks The NovoCyte flow cytometer needs to be shipped Abnormal warning about the liquid level in the reagent containers is observed The optical filters and mirrors are dirty 2 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 failed, 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 Debubble from the NovoExpress software to execute the debubble procedure. 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. 23

24 Unscheduled Maintenance Unclog the Sample Injection Probe (SIP) If the problem still exists after Backflush, follow the procedure below to manually clear the clog inside the SIP. Procedure 1 Turn off the power of the instrument. 2 Open the top cover of the instrument. 2 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 Maintenance. Figure 2-1 Remove the SIP from the Instrument The sample injection probe is fragile and may be damaged if handled improperly. Be careful when handling the sample injection probe to avoid deforming or bending it. 4 Connect the SIP with the 1/16 ID tubing provided in the NovoCyte Sample Injection Probe Unclog Kit. Connect 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 multiple times to flush the SIP with ACEA NovoClean solution. 6 Disconnect the SIP and insert it into a beaker containing ACEA NovoClean solution (1X) and soak for 10 minutes. uuu 24

25 Unscheduled Maintenance Clean the External Surface of the Instrument 7 Immerse the SIP into a beaker containing ACEA NovoFlow sheath fluid and soak for 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. 2.3 Clean the External Surface of the Instrument Keep the external surface of the instrument clean using the following method. 2 Materials ACEA NovoRinse solution (1X) Deionized (DI) water Clean soft cloth 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). 3 Wipe the surface again using a dry clean soft cloth. 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 Clean soft cloth 25

26 Unscheduled Maintenance Clean the Sample Injection Probe (SIP) ACEA NovoClean solution (1X) Deionized (DI) water Procedure 1 Click Instrument Shut Down on the NovoExpress software. 2 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. 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 directly contacts the biological samples which may be biohazardous. Wear gloves when handling 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. uuu 26

27 Unscheduled Maintenance Decontamination 5 Click OK button after cleaning. (Figure 2-5). 2 Figure 2-5 Message in NovoExpress Software to Conduct the SIP Cleaning Procedure 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 automatically. Figure 2-6 Message in NovoExpress Software to Conduct the SIP Cleaning Procedure 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 occurrence of bacterial contamination in the NovoCyte fluidics system. A Version 1 Decontamination Kit (Catalog No ) is required to run this procedure. The Decontamination function is only available by logging in as account with the Decontaminate Instrument privilege. 27

28 Unscheduled Maintenance Clean Fluidic System before Long-Term Storage Procedure 1 Click Instrument Fluidics Maintenance Decontamination from the NovoExpress software. 2 NovoExpress software will prompt a dialog box to guide you conducting the decontaminating 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. 2 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 cytometer for more than two weeks, a thorough 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 deionized water. 2 Click Instrument Fluidics Maintenance Priming from the NovoExpress software. Repeat the procedure two times. 3 Press the power switch on the front panel of the NovoCyte instrument to initiate the automatic shutdown cleaning process. 4 After the shutdown cleaning process is completed, the instrument will power off automatically. 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. When using the system after long-term storage, follow the procedure below to prepare the system. 28

29 Unscheduled Maintenance Purge Fluidic System before Shipment 1 Sterilize the reagent containers following the procedure described in Sterilize Instrument Reagent Containers of Section 1.1 Preventative Maintenance. 2 Add 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. 4 Click Instrument Fluidics Maintenance Priming from the NovoExpress software. Repeat the procedure two times. 5 The NovoCyte instrument is then ready to use. If NovoCyte instrument has not been used for more than two months, it is recommended to replace the fluidic system consumables before use. Refer to Section 1.2 Replacing Fluidic System Consumables for detailed procedure 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 properly for shipment. 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. 29

30 Unscheduled Maintenance 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 recalibrate 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 recalibrate the fluidics station. 2 Calibrate Fluidics Station function is only available when login as ccount 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 that the fluidics station calibration is successful. 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 30

31 2.9 Clean the Optical Filters and Mirrors Unscheduled Maintenance 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, the optical filters and mirrors may acquire dust or debris if unplugged from the slot for rearrangement or replacement. The following method describes how to clean the optical filters or dichroic mirrors. Keeping the optical filters and mirrors surface clean is very important for data quality. 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 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 filters. Materials Analytical grade ethanol Compressed air or a bulb blower Lint-free cotton swabs Lint-free wipe or lens cleaning tissue 2 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 new wipe and swab and repeat steps 2-3 again. 5 Repeat steps 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. 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. 31

32 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. 3 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 messages. Please contact ACEA Technical Support if issues are not resolved after performing the recommended 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 Zero or low number of events detected QC particles are expired or not stored appropriately Flow cell is dirty Air bubbles exist in the flow chamber Threshold is set too high Sample concentration is too low Sample is not mixed well NovoFlow container is empty NovoFlow container is not correctly connected to the instrument and the NovoFlow solution does not flow through the tubing Prepare a new tube of QC particles and mix them well to run the QC test again. Perform the maintenance functions from the NovoExpress software: Unclog, Priming, Extensive Rinse, and Extensive Clean. Execute Debubble and Priming function from NovoExpress Software. Lower the threshold setting and re-collect the sample. Redo sample preparation using an appropriate method to concentrate the sample. Mix the sample well to suspend the cells. Refill the NovoFlow container following the procedures described in Section Add Instrument Reagents in NovoCyte Flow Cytometer Operator s Guide. Reconnect the NovoFlow quick coupling connectors to ensure the NovoFlow container is correctly connected to the instrument. Execute Prime function from the Novo- Express Software. uuu 32

33 Troubleshooting Observation Possible Causes Recommended Solutions 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 NovoExpress Software. Sample tubing is leaking Clean the leaked fluid with wipes and properly reconnect the sample tubing. High CV Air bubbles exist in the Sampling Pump Air bubbles exist in the flow chamber Execute Debubble and Priming function from NovoExpress Software. Execute the Debubble and Priming function from NovoExpress Software. 3 Sampling tubing is clogged Perform maintenance functions from NovoExpress software: Unclog, Priming, Extensive 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 preparation 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 display 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-collect the sample. Fluidic system tubing is dirty Execute Rinse function from NovoExpress Software. Execute Decontamination function from NovoExpress Software. High event rate The threshold is set too low Increase the threshold setting and re-collect the sample. Sample concentration is too high Re-prepare the sample to appropriate concentration, or dilute the sample. Sample flow rate is too high Reduce the sample flow rate. NovoExpress displays Instrument not connected even though the instrument is powered up Workstation is in hibernation causing the interruption of communication between the NovoCyte instrument and workstation Restart NovoExpress Software. uuu 33

34 Troubleshooting Observation Possible Causes Recommended Solutions NovoExpress displays Instrument not connected even though the instrument is powered up Improper connection between the NovoCyte instrument and workstation Reconnect the USB cable between the instrument and the workstation. More than one NovoExpress instances are running. Only run one instance of NovoExpress software. Unstable Signal or gap in the data plot Air bubbles exist in the flow chamber Execute the Debubble and Priming function from NovoExpress Software. 3 Sample tube does not fit the sample tube holder Sample Injection Probe is not properly aligned with the wells or tubes Wrong type of sample tube is used Plate or tube rack is not loaded correctly. Use mm flow tube or 1.5 ml Eppendorf tube. Load the plate or tube rack with the correct 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 deformed or kinked Waste filter is clogged Reconnect the waste tubing to ensure it is properly connected. Move the instrument forward as needed to ensure sufficient space between the instrument and wall. Refer to Section Installation in NovoCyte Flow Cytometer Operator s Guide for more information on properly installing NovoCyte instrument. Replace the waste filter following the procedures described in Section 1.2 Replacing Fluidic System Consumables in this guide. Leakage from Sampling Pump Leakage from other components of the instrument Incorrect tubing connection to the sampling pump Loose fluidic components connection or or blocked sheath in-line filter Reconnect the tubing to the sampling pump. 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. Table 3-1 Troubleshooting Guide for NovoCyte Instrument 34

35 Troubleshooting Message IDs Software Messages Possible Causes Recommended Solutions 0x0001 Collision of SIP The movement of SIP is blocked by some obstacles Incorrect plate is selected in plate manager Incorrect positioning of plate in NovoSampler (Pro) Dirty SIP or SIP wash apparatus 0x0002 Running out of NovoFlow NovoFlow solution is not sufficient to continue to run any samples Fluidic station is not working properly Locate and clear the obstacles. Click the OK button in the prompted dialog box or wait 10 seconds for automatic error handling. The instrument will move the SIP to the home position. This procedure will take about 3 minutes. Select the correct plate in the plate manager window. Position the plate on the shaker correctly. Ensure the plate is seated flat on the stage inside the clamps. Re-calibrate NovoSampler Pro. Clean the SIP or SIP wash apparatus following the procedures described in Section 1.1 Preventative Maintenance in this guide. Refill the NovoFlow container following the procedures described in Section Add Instrument Reagents in NovoCyte Flow Cytometer Operator s Guide. Replace the fluidic station. 3 0x0003 Running out of Novo- Rinse NovoRinse solution is not sufficient to continue to run any samples Refill the NovoRinse container following the procedures described in Section Add Instrument Reagents in NovoCyte Flow Cytometer Operator s Guide. Fluidic station is not working properly Replace the fluidic station. 0x0004 Running out of Novo- Clean NovoClean solution is not sufficient to continue to run any samples Refill the NovoClean container following the procedures described in Section Add Instrument Reagents in NovoCyte Flow Cytometer Operator s Guide. Fluidic station is not working properly Replace the fluidic station. 0x0005 Waste container is full Waste container is too full to continue to run any samples Fluidic station is not working properly Empty the waste container following the procedures described in Section Empty Waste in NovoCyte Flow Cytometer Operator s Guide. Replace the fluidic station. uuu 35

36 Troubleshooting Message IDs Software Messages Possible Causes Recommended Solutions 0x0006 System voltage is out of range System Error Restart the NovoCyte instrument. 0x0007 System electric current is out of range System Error Restart the NovoCyte instrument. 0x0008 Firmware configuration error System Error Restart the NovoCyte instrument. 3 0x0009 0x000A 0x000B 0x000C 0x000D 0x000E laser self test error laser is not connected Specified laser is not functioning properly Specified laser is not detected The instrument will automatically reset the laser and run a laser self-test. It takes approximately 5 to 10 minutes. Restart the NovoCyte instrument. 0x000F NovoSampler communication 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 communicating 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) calibration failed NovoSampler (Pro) is not installed properly Re-install and calibrate the NovoSampler (Pro) following the procedures described in Section 1.2 Installation in NovoSampler Operator s Guide or NovoSampler Pro Operator s Guide. The NovoSampler (Pro) cover was opened during calibration. 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 NovoSampler Pro Calibrate in NovoExpress Software to re-calibrate the NovoSampler (Pro). uuu 36

37 Troubleshooting Message IDs Software Messages Possible Causes Recommended Solutions 0x0012 The movement of plate is out of range NovoSampler (Pro) is not installed properly Re-install and calibrate the NovoSampler (Pro) following the procedures described in Section 1.2 Installation in NovoSampler Operator s Guide or NovoSampler Pro Operator s Guide. 0x0013 Cover of NovoSampler (Pro) is opened during moving plate The cover of NovoSampler (Pro) is opened during moving plate 0x0014 Pressure is out of limit Waste container is not correctly connected to the instrument Sheath in-line filter is clogged Flow cell is clogged Close the NovoSampler (Pro) cover. NovoSampler (Pro) will automatically reset and be ready for operation. Check the quick coupling connectors to ensure the waste container is correctly connected to the instrument. Replace the sheath in-line filter following the procedures described in Section 1.2 Replacing Fluidic System Consumables in this guide. Perform maintenance procedures from NovoExpress software, Unclog, Priming, Extensive Clean, Extensive rinse. 3 0x0015 NovoSampler firmware error NovoSampler (Pro) firmware is not working properly Re-install or upgrade the NovoSampler (Pro) firmware. 0x0016 0x0017 0x0018 laser does not emit Specified laser is not connected properly or laser is not working properly Reconnect the laser properly. Restart the instrument. 0x0019 0x001A 0x001B laser communication 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 damaged 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 Bad connection or optocoupler is damaged Restart the instrument. 0x001E The SIP module needs to be upgraded to support NovoSampler Pro SIP module is not compatible with connected NovoSampler Pro Contact ACEA technical support for service. uuu 37

38 Troubleshooting Message IDs Software Messages Possible Causes Recommended Solutions 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 instrument 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. 3 0x0021 0x0022 Sheath fluid pump reset failed Resetting NovoSampler Pro to zero position failed Bad connection or optocoupler is damaged NovoSampler Pro is not working properly. Restart the instrument. Click Reset. Restart the NovoSampler Pro. 0x0023 Resetting NovoSampler Pro to home position failed NovoSampler Pro is not working properly. Click Reset. Restart the NovoSampler Pro. 0x0100 Instrument cover opened Instrument cover is opened or not tightly closed 0x0101 NovoFlow running low NovoFlow solution is below the pre-set volume limit Fluidic station not working properly 0x0102 NovoRinse running low NovoRinse solution is below the pre-set volume limit Fluidic station is not working properly 0x0103 NovoClean running low NovoClean solution is below the pre-set volume limit Fluidic station is not working properly Close the instrument cover. Refill the NovoFlow container following the procedures described in Section Add Instrument Reagents in NovoCyte Flow Cytometer Operator s Guide. Replace the fluidic station. Refill the NovoRinse container following the procedures described in Section Add Instrument Reagents in NovoCyte Flow Cytometer Operator s Guide. Replace fluidic station. Refill the NovoClean container following the procedures described in Section Add Instrument Reagents in NovoCyte Flow Cytometer Operator s Guide. Replace fluidic station. 0x0104 Waste container is close to full Waste is above the pre-set volume limit Empty the waste container following the procedures described in Section Empty Waste in NovoCyte Flow Cytometer Operator s Guide. Fluidic station is not working properly Replace fluidic station. uuu 38

39 Troubleshooting Message IDs Software Messages Possible Causes Recommended Solutions 0x0105 Cover of NovoSampler (Pro) is opened Cover of NovoSampler (Pro) is opened Close the cover. 0x0106 NovoSampler (Pro) is disconnected when powered up The NovoSampler (Pro) is disconnected 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 instrument is not properly connected Power down the instrument. Reconnect the cable between the fluidics station and the NovoCyte instrument. Power up the instrument. 3 0x010A 0x010B 0x010C 0x010D liquid level sensor failure Specified liquid level sensor in the fluidics station is not working properly Reconnect the fluidics station cable. Restart the NovoCyte instrument. Replace the fluidics station. 0x010F Sheath filter is clogged. Please replace the sheath filter and run the Priming procedure 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. Communication error (code: xx, xx). Please restart NovoCyte and NovoExpress USB cable between the NovoCyte instrument and workstation is not connected properly Reconnect the USB cable between the NovoCyte instrument and the workstation. Restart the instrument and NovoExpress software. Table 3-2 NovoExpress Prompted Error Messages and Message IDs 39

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42 Published by ACEA Biosciences, Inc Mesa Ridge Road Ste 100, San Diego, CA U.S.A. (866) ACEA Biosciences, Inc. All rights reserved. ID150059