GeminiBio ORFLO Moxi V User manual

For Research Use Only. Not For Use In Diagnostic or Therapeutic Procedures.
a subsidiary of
USER GUIDE

Document # MXV.SYS.004
Effective: 30JUL2021
Revision # 2.12
Page 2 of 108
Title: Moxi V – User Guide
This document is provided to customers who have purchased Orflo Technologies, LLC (“Orflo”, a subsidiary of
Gemini BioProducts, LLC) equipment, software, reagents and consumables to use in the operation of such Orflo
equipment, software, reagents, and consumables. This document is copyright protected and any reproduction of this
document in whole or any part is strictly prohibited, except as Orflo may authorize in writing.
Limitations: The Moxi V™ is intended for research use only. NOT for use in diagnostic procedures. It is not
intended for use in medical diagnostic or therapeutic procedures, and it has not been approved or cleared by the
Unites States Food and Drug Administration for such uses.
Restrictions of Use: Software that may be described in this document is furnished under a license grant. Customer
shall not, nor shall it permit any third party to: (i) use, copy, modify, translate or adapt the Software or Documentation
other than as expressly permitted by the Terms of Sale, which can be found here
(http://www.orflo.com/v/vspfiles/documents/termsandconditions.pdf); (ii) rent, lease, share, disclose or transfer the
Software, provide it on a subscription basis, or use it in connection with a service bureau or other configuration
whereby any third party may use the Software, (iii) create any derivative works of or based on the Software or
Documentation; (iv) reverse-engineer, decompile, disassemble or otherwise attempt to discern the source code of the
Software; (v) remove, relocate, alter or obscure any trademark, copyright or other proprietary or restrictive marking or
legend on the Software or Documentation or any copies thereof; or (vi) use, provide or disclose the Software in
violation of any applicable laws, orders or regulations.
Disclaimer: The information in this document is subject to change without notice and should not be construed as a
commitment by Orflo. Neither Orflo nor any of its affiliated corporations assumes responsibility for any errors that
may appear in this document. This manual is believed to be complete and accurate at the time of publication. In no
event shall Orflo or any of its affiliated corporations be liable for incidental or consequential damages in connection
with or arising from the use of this manual.
Trademarks: Portions of this document may make reference to other manufacturers and/or their products, which
may contain parts whose names are registered as trademarks and/or function as trademarks of their respective
owners. Any such usage is intended only to designate those manufacturer’s products as supplied by Orflo for
incorporation into its equipment and does not imply any right or license to use or permit others to use such
manufacturer’ and or their product names as trademarks.
© 2021 Gemini BioProducts, LLC All rights reserved.
Rev. 2.12, 06/2021
Orflo Technologies and Moxi V™ are trademarks of Gemini BioProducts, LLC, registered in the U.S. and other
countries.
Windows, Windows XP, Windows Vista, Windows 7, and Windows 8, Windows 10 are trademarks of Microsoft
Corporation, registered in the U.S. and other countries.
Mac and Mac OS are trademarks of Apple Corporation, registered in the U.S. and other countries.
FlowJo is a trademark of Treestar, Inc.
Flowmi™ is a trademark of Bel-Art.
Bluetooth® and the Bluetooth logos are registered trademarks of Bluetooth SIG, Inc., registered in the U.S. and other
countries.
Accutase® and Accumax® are registered trademarks of Innovative Cell Technologies.
TÜV SÜD and the TÜV SÜD symbol are registered trademarks of TÜV SÜD Aktiengesellschaft.

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Revision # 2.12
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Title: Moxi V – User Guide
Contents
INTRODUCTION ................................................................................................................................................... 1
ABOUT THE USER GUIDE ...................................................................................................................................... 1
CONVENTIONS USED IN THE USER GUIDE ............................................................................................................ 2
SAFETY PRECAUTIONS ......................................................................................................................................... 2
ELECTRICAL SAFETY ........................................................................................................................................................ 2
LASER SAFETY ............................................................................................................................................................... 3
BIOLOGICAL SAFETY ....................................................................................................................................................... 3
MOXI V™ SYSTEM OVERVIEW .............................................................................................................................. 5
MOXI VTM ACCESSORIES ....................................................................................................................................... 7
GETTING STARTED ............................................................................................................................................... 8
GENERAL GUIDELINES .......................................................................................................................................... 8
ADDITIONAL MATERIALS REQUIRED .................................................................................................................... 9
USING THE MOXI VTM .......................................................................................................................................... 10
HOME (START) SCREEN ................................................................................................................................................. 11
SETTINGS ................................................................................................................................................................... 12
Fluorescence Gain ............................................................................................................................................... 12
Date and Time .................................................................................................................................................... 12
Auto Noise-Find .................................................................................................................................................. 13
Auto Gate settings .............................................................................................................................................. 13
Idle Shutdown Timer ........................................................................................................................................... 13
Password Expiration ........................................................................................................................................... 13
Secure Mode ....................................................................................................................................................... 13
SAMPLE PREPARATION CONSIDERATIONS ......................................................................................................................... 13
RUNNING A TEST ......................................................................................................................................................... 15
Instructions – Cell Count (Size + Viability), PBMC Check, CAR T Expansion Tests ............................................... 15
• Inserting a Cassette: ................................................................................................................................................ 15
• System Calibration: ................................................................................................................................................. 15
• Loading a sample: .................................................................................................................................................... 16
“Cell QC” Test ..................................................................................................................................................... 18
Cell QC – Single Run .......................................................................................................................................................... 19
Cell QC – Start Batch ......................................................................................................................................................... 20
Cell QC – Initial Test Output (both Single Run and Batch) ................................................................................................ 21
Cell QC – Test Output – Continuing Batch Testing ............................................................................................................ 23
Cell QC – Batch Run – Saved Data and .CSV Summary File ............................................................................................... 24
Cell QC – “Add to Existing Batch” ...................................................................................................................................... 25

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TEST RESULTS ............................................................................................................................................................. 25
Recorded Channels ............................................................................................................................................. 25
Data Display Formats – General Output Types .................................................................................................. 26
Fluorescence vs Size Scatter Plot ...................................................................................................................................... 27
Histograms ........................................................................................................................................................................ 29
Reading Raw Data Output (Conc., Size, MFI, and %) .......................................................................................... 31
Fluorescence vs. size scatter (dot) plot ............................................................................................................................. 31
Single Channel Histogram Plot .......................................................................................................................................... 32
Simplified Output and Auto-Gating Overview .................................................................................................................. 32
“PBMC Check" Output ........................................................................................................................................ 33
“CAR-T Expansion" Output ................................................................................................................................. 35
“Cell QC” – Output (both Single Run and Batch) ................................................................................................ 36
ANALYZING/GATING THE DATA ...................................................................................................................................... 38
(General) Recommended Post-Test Workflow ................................................................................................... 39
Re-scaling the X axis (size range) ........................................................................................................................ 39
Gating the data .................................................................................................................................................. 40
Overview ........................................................................................................................................................................... 40
Size Gating of Scatter (Dot) Plots ...................................................................................................................................... 42
Fluorescent Gating of Scatter (Dot) Plots ......................................................................................................................... 43
Noise Gating of Scatter (Dot) Plots ................................................................................................................................... 44
Store and Recall of Gate Settings ...................................................................................................................................... 45
Histogram Overlays on Fluorescence vs Size Scatter Plots ............................................................................................... 46
“Size Histogram” Assay ..................................................................................................................................................... 46
Changing Display Axis for Tests (Fluorescence vs Size, Histogram Only Views) ................................................. 49
FILE/TEST OPERATIONS ................................................................................................................................................ 51
Test Naming and Renaming Tests ...................................................................................................................... 51
Saving and Deleting Data/Tests ......................................................................................................................... 51
Exporting Data Screenshots ................................................................................................................................ 52
Editing Saved Tests ............................................................................................................................................. 53
Compare/Overlay Saved Tests ............................................................................................................................ 55
EXPORTING DATA – CONNECTING VIA USB (USB ON-THE-GO) ............................................................................................ 58
Connect the Moxi VTM to a Computer ................................................................................................................. 58
Put the Moxi VTM in USB Connect mode ............................................................................................................. 59
Safely disconnect Moxi VTM from the Computer ................................................................................................. 61
MOXI VTM HELP AND SYSTEM INFORMATION .................................................................................................................... 61
MOXI VTM APPS .......................................................................................................................................................... 62
21 CFR PART 11 OPERATION (“SECURE MODE”) .................................................................................................. 64
OVERVIEW ................................................................................................................................................................. 64
ACTIVATING SECURE MODE (21 CFR, PART 11 COMPLIANT MODE) ...................................................................................... 64
SECURE MODE – KEY SETTINGS ...................................................................................................................................... 66
Idle Shutdown Timer ........................................................................................................................................... 67

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Password Expiration ........................................................................................................................................... 67
Secure Mode ....................................................................................................................................................... 67
SYSTEM LOGIN ............................................................................................................................................................ 67
USER ADMINISTRATION ................................................................................................................................................ 68
Adding a User ..................................................................................................................................................... 68
User Roles ........................................................................................................................................................... 70
Modifying Users .................................................................................................................................................. 71
Deleting Users .................................................................................................................................................... 72
RUNNING TESTS – SECURE MODE ................................................................................................................................... 73
Saving Data ........................................................................................................................................................ 73
Electronic Signature .......................................................................................................................................................... 73
File Save Formats .............................................................................................................................................................. 75
FCS data ....................................................................................................................................................................... 75
BMP File ....................................................................................................................................................................... 75
REVIEWING DATA – SECURE MODE ................................................................................................................................. 76
SYSTEM EVENT LOGGING – SECURE MODE ....................................................................................................................... 79
Log File Naming .................................................................................................................................................. 80
Logged System Events ........................................................................................................................................ 80
BACKING UP / TRANSFERRING DATA – SECURE MODE ........................................................................................................ 81
Test-Files Directory ............................................................................................................................................. 81
Users Directory ................................................................................................................................................... 81
INSTRUMENT FIRMWARE AND SOFTWARE UPGRADES ....................................................................................... 82
UPGRADE INSTRUCTIONS: .............................................................................................................................................. 82
DATA (µSD) DISK FORMAT (STRONGLY RECOMMENDED) ....................................................................................... 83
MOXI V – TRAY INSERT REPLACEMENT PROCEDURE ........................................................................................... 86
BACKGROUND ............................................................................................................................................................. 86
ORDERING INFORMATION .............................................................................................................................................. 86
REMOVING AN EXISTING INSERT ...................................................................................................................................... 86
INSTALLING AN INSERT .................................................................................................................................................. 87
TROUBLESHOOTING ........................................................................................................................................... 89
MAINTENANCE AND STORAGE ........................................................................................................................... 94
SPECIFICATIONS FOR THE MOXI V™ AND CASSETTES ........................................................................................... 97
ORDERING INFORMATION .................................................................................................................................. 99
TECHNICAL AND CUSTOMER SERVICE ................................................................................................................. 99
WARRANTY ...................................................................................................................................................... 100
APPLICABLE PATENTS ....................................................................................................................................... 101

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Title: Moxi V – User Guide
Moxi V™ User Guide
Page 1
Introduction
The Moxi V™ from Orflo Technologies (a subsidiary of Gemini BioProducts, LLC) is a benchtop
instrument that utilizes a disposable thin-film cassette and a combination of a 532 nm laser, one
PIN diode (with a 561nm/LP filter), Coulter Principle-based cell measurements, and on-board
software to provide easy-to-run applications and data analysis. The instrument relies on
disposable cassettes for sample handling, which alleviates the need for flow cell cleaning and
fluidics maintenance. The instrument is small enough to be portable between the lab bench and
the hood. A touchscreen interface and simplified GUI allows for ease of operation without
extensive training.
The Moxi V enables highly accurate cell counts, cell sizing, quantitative assessments of cell
viability. Data acquisition occurs within approximately 5-10 seconds per test. The instrument
can store up to 4Gb of data. Data is provided in the industry-standard Flow Cytometry Standard
(FCS v3.1) format as well as user-generated bitmap screenshots. Data can be downloaded to a
PC or Mac via USB connectivity.
IMPORTANT NOTE:
The Moxi V™ is intended for research use only. Not for use in diagnostic procedures. It
is not intended for use in medical diagnostic or therapeutic procedures, and it has not been
approved or cleared by the United States Food and Drug Administration for such uses.
About the User Guide
The Moxi V™ User Guide provides detailed information for operating, maintaining, and
troubleshooting the Moxi V, including the system, the cassettes, and application workflows.
This includes information on how to analyze test results using the onboard analysis features of
the software.
The user guide does not include instructions for analyzing the provided/output flow cytometry
standard (FCS, version 3.1) files offline in third party software.
Similarly, the user guide does not include detailed protocols for preparing samples prior to
running them on the instrument. The results of the assays are dependent upon the proper use
of reagents and the instrument. Please refer the Protocols section
(http://www.orflo.com/v/vspfiles/pages/protocols.html) of the Orflo website for more details on
sample preparation.

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Title: Moxi V – User Guide
Moxi V™ User Guide
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To update the Moxi V firmware to the latest version, please visit http://www.orflo.com, and
register your instrument by entering a valid email address and serial number. The registration
key provided after registering will allow users access future firmware updates.
Conventions Used in the User Guide
Within this manual, the following conventions are used to call out certain points, and also to
reminder users of areas of precaution for safety reasons.
WARNING Alerts you to a situation that may cause injury to the user.
CAUTION Alerts you to a situation that may cause damage to the system, loss of
data, or incorrect results.
NOTE Indicates additional related information that may be helpful to the user.
Safety Precautions
The Moxi V™ has integrated safety features that are designed for the protection of the user. THE
INSTRUMENT SHOULD BE USED ONLY BY PROPERLY QUALIFIED AND TRAINED
LABORATORY PERSONNEL. IT SHOULD BE USED ONLY AS DIRECTED IN THIS GUIDE.
PLEASE REVIEW AND UNDERSTAND ALL SAFETY INSTRUCTIONS BELOW, AND AS
INDICATED THROUGHOUT THE USER GUIDE, BEFORE OPERATING THE SYSTEM.
Electrical Safety
WARNING To avoid the danger of electric shock, including the possibility of permanent
injury or death:
• Prior to use, verify that the USB cable and USB charging adapter are plugged
securely into a properly grounded AC power outlet. Verify that the connection
between the USB cable and the instrument is secure and the AC power in your
location is within the specifications for the instrument.
• Do not immerse the instrument, USB cable, or USB power adapter in liquid or allow
liquid to enter the instrument.
• Do not attempt to disassemble or service the Moxi V. Aside from swapping of filters
and replacement of cassette tray inserts, the instrument has no user serviceable

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parts. All service must be performed by ORFLO, or an approved ORFLO
vendor.
Laser Safety
WARNING The Moxi V is a Class 1 laser product in accordance with IEC 60825-1:2007.
The system contains a Class IIIb laser that operates at 532 nm with a
maximum output power of 50 mW. Direct exposure to laser radiation, even
for a fraction of a second, can be harmful to skin, eyes, or other body parts,
including posing a risk of burns and permanent vision impairment or
blindness. The following guidelines should be followed to avoid exposure:
• An interlock on the door prevents the laser from operating when the door is open.
Do not attempt to defeat the interlock and run a test with the door open.
• Do not operate the Moxi V if there is any damage to the instrument’s enclosure.
• Do not attempt to disassemble or service the Moxi V to service or adjust the laser.
The instrument has no user serviceable parts, and attempts to adjust or service the
laser may result in hazardous radiation exposure. All service must be performed
by ORFLO, or an approved ORFLO vendor.
Biological Safety
WARNING Biological samples have the potential to transmit harmful or fatal disease.
The following guidelines should be followed to minimize the risk of exposure
to biohazardous materials.
• Handle all biological samples as if they are capable of transmitting harmful or fatal
infections.
• Wear appropriate personal protective equipment (PPE) including clothing, eyewear,
and gloves.
• Do not pipette by mouth.
• Dispose of biological waste in accordance with all applicable local, state, and
federal regulations.

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Title: Moxi V – User Guide
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Moxi V™ System Overview
The Moxi V™ Kit includes the Moxi V system, USB Cable (mini USB B to USB A 2.0), USB
Power Adapter, one pack of 25 Type S+ (MXC030) Cassettes, a spare cassette tray insert, and
a Quick Start Reference Guide. The following figure shows the main visible components of the
Moxi V and the table provides a description of each component.