Related Manuals for GeminiBio ORFLO Moxi V
Summary of Contents for GeminiBio ORFLO Moxi V
- Page 1 a subsidiary of USER GUIDE For Research Use Only. Not For Use In Diagnostic or Therapeutic Procedures.
- Page 2 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.
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Page 3: Table Of Contents
Contents INTRODUCTION ..............................1 ABOUT THE USER GUIDE ............................1 CONVENTIONS USED IN THE USER GUIDE ......................2 SAFETY PRECAUTIONS ............................2 ................................ 2 LECTRICAL AFETY ................................2 ASER AFETY ............................... 3 IOLOGICAL AFETY MOXI V™ SYSTEM OVERVIEW ..........................4 MOXI V ACCESSORIES ............................ - Page 4 Simplified Output and Auto-Gating Overview ........................25 “PBMC Check" Output ............................26 “CAR-T Expansion" Output ..........................27 “Cell QC” – Output (both Single Run and Batch) ....................29 ............................30 NALYZING ATING THE (General) Recommended Post-Test Workflow ....................31 Re-scaling the X axis (size range) ........................31 Gating the data ..............................
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Page 5: Introduction
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. -
Page 6: Conventions Used In The User Guide
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. -
Page 7: Biological Safety
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: •... -
Page 8: Moxi V™ System Overview
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. -
Page 9: Moxi V
Component Function Touch Screen Display Allows user to interface with the instrument by pressing icons and targets. Displays all information needed for operation and analysis of results. Power/Reset Button Turns instrument on and off and resets the unit when pressed and held for ~10 seconds. Cassette Tray Door The door is manually opened and closed by user to allow insertion of a cassette into cassette tray. -
Page 10: Getting Started
Getting Started The Moxi V™ is shipped in a condition ready for initial use; though we recommend checking for software updates and updating the Moxi V with the most current software version upon receipt. To begin, plug the USB Cable into the instrument at one end (mini-USB). Plug the opposite end of the USB Cable (standard USB) into the Power Adapter. -
Page 11: Additional Materials Required
For cell counting application, cells can be run without fluorochromes using any of the assays. Additional Materials Required • Cell or bead sample (diluted and dissociated, if necessary); 60 µL minimum. • Type S+ (MXC030, recommended for almost all assay) or MF-M (MXC010) Cassettes •... -
Page 12: Using The Moxi V
Using the Moxi V Press the Power Button to turn on the instrument. The Home Screen will become available. The image below identifies the key items on the Home Screen. Home (Start) Screen Battery/ Charging Indicator Date/ Time Indicator Assays /Applications Help Instructions... -
Page 13: Settings
Settings Date and Time Set the date and time by pressing the Settings icon on the main menu of the instrument. Then touch the Date or Time field. Use the up/down arrow keys to change the values. Passcode Lock The Moxi V can be setup to require a password login to use the instrument and access stored data. -
Page 14: Fluorescence Gain
it on (See images below). Select Done. The system will present a keyboard to enter the four- digit numeric passcode. On entering the passcode, the user will be required to “Re-enter Passcode” to prevent accidental entry of a wrong code. Upon successful re-entry of the passcode, the system will display a “lock enabled”... - Page 15 1. Solution Conductivity – As the system utilizes an electrical measurement for counting cells, sizing cells, and metering the fluid volume, cells need to be suspended in a conductive media. The target solution conductivity would be 0.9% salt solution (e.g. PBS or equivalent).
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Page 16: Running A Test
Running a Test Instructions – Cell Count (Size + Viability), PBMC Check, CAR T Expansion Tests • Please follow sample preparation guidelines above (“Sample Preparation Considerations”) in preparing the sample. Note: The minimum volume required for a test is 60 µL. • Turn the instrument on by pressing the power button and the Home screen will be displayed. -
Page 17: Loading A Sample
• Loading a sample: System messages at the top of the screen prompt the user to “Open the Door” (image below/left) and pipette a 60 – 75 µL sample (“Enter the Sample”, image below/middle) into the fill port of the cassette (either test 1 or test 2, depending on which end of the cassette was inserted into the instrument) and close the Door. -
Page 18: "Cell Qc" Test
• It is best to place the pipette facing straight into the system (not angled left or right) and a ~45 degree angle (up/down, see image below) • Dispense the fluid in one, smooth motion as you would normally during pipetting. The system does apply a slight vacuum to pull the fluid in, but it is not recommended to try to “time the load”... -
Page 19: Cell Qc - Single Run
appending to an existing batch run). The selected option is display in orange. The other options in green. Cell QC – Single Run By default, the system sets the “File Option” as a “Single Run” (single test, see image above/middle). Below the file option selection, there are three test parameters that the user can set: 1.) “File Name”... -
Page 20: Cell Qc - Start Batch
Cell QC – Start Batch Selecting the “Start Batch” file option will prompt a warning that this mode is only meant for samples of the same cell type (see image above/middle). The reason for this is that, following the first test (which will be auto-gated by the system, with the option to manually adjust the gates), the subsequent tests will be run with the same gate settings as the first. -
Page 21: Cell Qc - Initial Test Output (Both Single Run And Batch)
test/cassette side, the system will perform a spiral find and laser sweep to find align the laser ideally to the cassette detection zone (see laser/cassette alignment section). Cell QC – Initial Test Output (both Single Run and Batch) An example of the Cell QC output is shown above. Initially (during and right after acquisition) the user is shown the raw scatter plot data (image above/left) and given the option (by touching the orange rectangle) to manually adjust the system’s auto-gate placement. -
Page 22: Cell Qc - Test Output - Continuing Batch Testing
• The key information, dilution-adjusted concentration and viability, are highlighted in the white box. • The “Debris %” is calculated as the particle counts in the range from 3µm (minimum detectable particle size) to the left size gate (start of the cell region). •... -
Page 23: Cell Qc - Batch Run - Saved Data And .Csv Summary File
Following completion of a batch, the user can exit the batch processing by touching the green “No” icon (image above/middle) to exit back to the home screen. Cell QC – Batch Run – Saved Data and .CSV Summary File The batch information is all contained in a sub-folder (under the “Test-Files” main folder) of the Moxi V µSD data disk. -
Page 24: Cell Qc - "Add To Existing Batch
Cell QC – “Add to Existing Batch” To append data to existing batch, the user can simply select the “Add to Existing Batch” icon (see above/left) at the first “Cell QC” configuration screen. The system will provide the standard warning (yellow pop-up, see above/left) that batch mode is meant for samples of the same type. Touch “Accept,”... - Page 25 on the system “Auto Gate Setting” setting (see “settings” section). Note: All test results are ultimately based on the gating of the raw data displays. Even with the simplified tests, the user always has the option to view/review the raw data and adjust the gates if needed. The raw data can be displayed in two main formats: 1.).
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Page 26: Fluorescence Vs Size Scatter Plot
Fluorescence vs Size Scatter Plot Test name Population % Test type Toggle between Total size-gated gates: Size (blue), (between blue gates) fluorescence (red), counts and noise (yellow) Left size marker Right size marker adjust (drag to move) adjust (drag to move) Fluorescence scale Fluorescence gate – (“Logicle”... -
Page 27: Histograms
Histograms Test type Test name Viability % (touch to change) (defined by red and blue gates) Swipe up/down in Histogram area to change y-axis scale Histogram bin counts Lower (negative/low Fluorescence) cell pop (green) – “Live Cells” Upper gated (above red Line defined in PMT vs Size view) pop results “Dead Cells... -
Page 28: Reading Raw Data Output (Conc., Size, Mfi, And %)
Reading Raw Data Output (Conc., Size, MFI, and %) The Moxi V™ provides particle concentration and precise sizing information for each gated region, total counts information, fluorescent percentages, and median fluorescent intensities (all where applicable based on the channels selected) for each test. Two example test view screens are shown above (left to right: Fluorescence vs. size scatter (dot) plot, Single Channel Histogram Display). -
Page 29: Simplified Output And Auto-Gating Overview
the green representing the fluorescence- (below the red line) points. Any areas of overlap in the histogram (between red and green populations) are shaded yellow. Note: The overlap is a natural consequence of converting the 2D scatter plot data to a 1D Histogram. The two black boxes below the histogram provide the subset concentration (cells/ml), mean particle diameter (Note: This is an “effective”... -
Page 30: "Pbmc Check" Output
“PBMC Check" Output The initial, simplified output of the PMBC test is shown above. This output is based on the system’s auto-gating of the PIN Diode fluorescence (Propidium Iodide, Viability) vs. Size scatter plot (more detail below) to identify the core PBMC population (by size) and determine the viability (by PI fluorescence). -
Page 31: "Car-T Expansion" Output
The Pin Diode (Propidium Iodide, viability) vs size scatter plot is display (above/middle). The placement of the size gates (blue markers, vertical lines, two top dashed red arrows in image above/middle) and fluorescence gate (horizontal red line, two lower dashed red arrows in image above/middle). - Page 32 The data scatter plots/histograms and associated system-set gates can be viewed and adjusted by the user. It is recommended, particularly for the first test in a sequence of experiments, that these gates be checked by the user. The scatter plot is accessed by touching the “Next” icon (image below/right).
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Page 33: Cell Qc" - Output (Both Single Run And Batch)
“Cell QC” – Output (both Single Run and Batch) An example of the Initial Cell QC “Results Summary” is shown in the image above/left. This initial screen is designed to provide a simple representation of all the cell/sample data based on the system auto-gating of the Pin Diode (Propidium Iodide, Viability) vs Size scatter plot (more detail below) to identify the core cell population (by size) and determine the viability (by PI fluorescence). -
Page 34: Analyzing/Gating The Data
The up/down arrows on the histogram can be used to resize the histogram y-axis (scaling). The “Print BMP” button will save a Bitmap (image, .bmp) file to the Moxi V data disk (in the main “Test-Files” folder for single runs or the batch folder for batch runs). That image will be an exact representation of the “Results Summary”... -
Page 35: (General) Recommended Post-Test Workflow
(General) Recommended Post-Test Workflow Adjust the Define Define the +/- Define the Read results size (x) population by & perform file axis to analysis adjusting the population operations optimize region by (e.g. export the size adjusting (fluorescence adjusting screenshot, resolution the size ) gate the red save, save (blue) -
Page 36: Gating The Data
Rescaling the axis results in reprocessing of the raw data to ensure that resolution is not lost during the rescale process (1000 histogram bins). This allows (and is recommended for) users to achieve finer resolution of smaller particle populations and potentially increased separation between particle sub-populations. -
Page 37: Size Gating Of Scatter (Dot) Plots
Size Gating of Scatter (Dot) Plots The results of all fluorescent tests (all tests except for the “Size Histogram” test) are initially displayed as scatter plots with the size (blue) gates enabled (see image above, left). Gates can be adjusted by touching and dragging the blue markers around the edges of the scatter plot (above). -
Page 38: Fluorescent Gating Of Scatter (Dot) Plots
cursor can even be dragged around the left edge of the scatter plot if needed. Events to the left of the left size gate and to the right of the right size gate are not included in the count totals or population percentages (e.g. -
Page 39: Noise Gating Of Scatter (Dot) Plots
to place the fluorescent gate at the top/tail of the noise cluster. Gate placement can also be faciliated by running control (negative or positive) tests, setting the gates (above or below the control, respectively), storing the gates locations (see section on storing/recalling gates), and then recalling the gates for the standard, labeled sample. -
Page 40: Store And Recall Of Gate Settings
Store and Recall of Gate Settings Moved Gates If the system auto-gating is not enabled for a particular test, the Moxi V™ gates are then “sticky” by default. “Sticky” means that each new test defaults to the gate settings (size and fluorescence) of the prior test. The instrument also allows the user to store up to five gate locations in memory for recall. -
Page 41: "Size Histogram" Assay
down for the size histogram and left-right for the fluorescent histogram. The image below/right shows the effect of making broad left-to-right swipes as well as top-to-bottom swipes in the scatter plot region of the middle image below. The result is that the fluorescence histogram scale is decreased (increasing histogram height) and the size histogram scale is increased (shrinking the size histogram). - Page 42 Swipe Swipe up/down up/down to Rescale to Rescale The Counts (Y axis) scale can be adjusted swiping your finger up or down in the white histogram area. For low concentration cell samples, it is important to decrease the counts scale (swipe from the bottom to top) in order to properly view and gate the population.
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Page 43: Changing Display Axis For Tests (Fluorescence Vs Size, Histogram Only Views)
Changing Display Axis for Tests (Fluorescence vs Size, Histogram Only Views) Y-axis Select X-axis Select FLUO FLUO The test output can be modified to display any recorded channel vs another channel or, individually, with a histogram-only view. The change the display axis, the X/Y button (image below, frame at the top/right) can be selected. When X/Y is pressed the available channels for each axis are displayed in the gray boxes at the top and to the right of the current scatter (image below, frame at top/middle) plot (or histogram). - Page 44 selected as the x-axis. Some examples are shown in the above figure (but not all possible permutations) including: • Pin Diode (561nm/LP) fluorescence histogram (image above, top/right frame). Displayed by selecting “Histo” at the top and “Fluo” at the right. Notes: o For fluorescence histograms, the binned data is defined by the size gated region in the corresponding fluorescence vs.
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Page 45: File/Test Operations
File/Test Operations Test Naming and Renaming Tests By default, tests are named with an automated prefix based on the test type (“Via” for Cell Counts (Size + Viability), “PBMC” for PBMC Check, “CAR” for CAR-T Expansion, “QC” for Cell QC, and “Size” for Size Histogram). Files within test types are auto-enumerated by the system (e.g. -
Page 46: Exporting Data Screenshots
All saved tests can be deleted as a batch by selecting the Delete All button in the Saved Data area of the instrument or by checking tests in the Saved Data area and selecting the Delete Selected button. Exporting Data Screenshots Exact data screenshots can be generated from the instrument by selecting File | Print | Export Screenshot (see image sequence below) from the test screen. - Page 47 Tests can be sorted by either Name, Date, or Type (FCS, BMP, etc.) by touching the corresponding icon in the “Sort by” field at the bottom left of the screen. Touching the same icon (e.g. Name) repeatedly reverses the sort order (ascending vs. descending). Test files and screenshot files can be opened by simply touching the green (or dark gray row).
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Page 48: Compare/Overlay Saved Tests
• Page up/down (ten files at a time) the list by sliding the up/down arrows to the right of the file list. Files can be selected for batch operations by touching the corresponding white checkbox (or deselected by touching the red “X” checkbox). Once files have been selected, the Compare/Overlay (Future functionality) and Delete Selected button functionality is activated (image below, top/right frame). - Page 49 Page 45 ™ Moxi V User Guide...
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Page 50: Exporting Data - Connecting Via Usb (Usb On-The-Go)
Exporting Data – Connecting Via USB (USB on-the-go) Each saved Moxi V™ test is saved in the Flow Cytometry Standard (FCS) v3.1 format (International society for Advancement of Cytometry, http://isac-net.org) and can correspondingly be opened by any commercially available FCS v3.1 compatible software package (e.g. -
Page 51: Put The Moxi Vtm In Usb Connect Mode
Put the Moxi V in USB Connect mode Touch the “USB Disk” icon (image above/left) Touch the “Connect Disk” icon (image above/middle) iii. Wait for the “Connected” icon to light up (image above/right). Notes: • It may take a couple of minutes to light up. • Once that disk is connected, the Moxi V disk will appear as an external drive on your computer. -
Page 52: Safely Disconnect Moxi Vtm From The Computer
Safely disconnect Moxi V from the Computer Safe-eject the Moxi V drive from the computer. • Windows: Click on the “Safely Remove Hardware” icon in the system tray (bottom right of Windows taskbar). Select the option to eject the Moxi V disk. • Mac: In the Mac Finder, click on the arrow to the right of the “Moxi V SD” external drive that corresponds to the Moxi V disk. -
Page 53: Moxi Vtm Apps
2. Press the Next icon to view the next visual instruction that demonstrates how to pipette a sample into the cassette. 3. Press the Next icon to proceed to the next instruction screen or press the Previous icon to return to the previous screen. The last screen of the help menu provides information on the version of the Moxi V™... -
Page 54: Instrument Firmware And Software Upgrades
• Restart – Reboots the unit • Off – powers the unit off. Instrument Firmware and Software Upgrades This User Guide describes OS version 2.10 of the Moxi V™ firmware. The firmware may be updated periodically. For information on the most up-to-date firmware, visit the ORFLO website at www.orflo.com. - Page 55 Page 51 ™ Moxi V User Guide...
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Page 56: Moxi V - Tray Insert Replacement Procedure
Moxi V – Tray insert replacement procedure Background The Moxi V has a tray design that allows users to address the occasional need to clean the system cassette tray of adhesive build-up. The adhesive build-up can eventually lead to increased resistance to the insertion and removal of cassettes from the system. When resistance to inserting/removing cassettes becomes prohibitive, users can simply remove a tray insert (that has the adhesive build-up), replacing it with a new insert (or an old one that has had the adhesive cleaned off). -
Page 57: Installing An Insert
2. Gently lift up on the center tab (red arrow in images below) of the tray insert while pulling out to slide the insert out. Pull insert all the way out. (Refer to images below) Installing an insert IMPORTANT: Make sure to unplug Moxi V USB/Power cable from system before starting this procedure. - Page 58 Tray Guides Insert Rails 3. Push the cassette insert into the system. Page 54 ™ Moxi V User Guide...
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Page 59: Troubleshooting
4. Once the insert is all the way in, try closing the cassette door. If insert is properly installed, door should close all the way. If the door doesn’t close, try to push the insert further in. Troubleshooting Symptom Cause Corrective Action Try charging the unit overnight (minimum, over a Battery gauge has reset or battery... - Page 60 Please check compatibility of the fluorophore with No Fluorescence Incorrect fluorophore was used. the system specifications (532nm laser excitation separation/detection and 561nm/LP filtered emission) Cell preparation protocol labeling step was No fluorophore present omitted. Make sure concentration of cell or bead sample is Concentration of cell/bead sample within recommended guidelines.
- Page 61 Overlapping size populations can obscure the Debris-filled sample or size- downward slopes of cell peaks. Often debris overlapping cell samples populations can obscure the left side of a cell (polydisperse samples). peak. Adjust pipette to ensure sufficient volume for the Insufficient fluid (<60 µL) test.
- Page 62 Test Error: Fluid was not Sample not loaded or volume too Make sure sample is loaded and volume is at Detected small least 60 µL. Adjust pipette to ensure sufficient volume for the Insufficient fluid (<60 µL) test. For improperly calibrated pipettes, this might require a setting >60 µL Test Error: Incomplete Test Make certain that the particle type is within the...
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Page 63: Maintenance And Storage
Maintenance and Storage Storage Store the Moxi V and Cassettes at room temperature in a dry environment. For best results, cassettes should be used within one year of purchase. Avoid prolonged exposure to ultraviolet light as it may degrade the touch screen display or discolor the external surfaces of the instrument. - Page 64 The Moxi V contains a 3.7 V, 7500mAh lithium ion battery. The battery is incorporated in the system only to buffer the power surges that occur during the activation of the laser and to ensure that a temporary unplugging of the system does not result in a power-off. The system is NOT intended to be operated under battery power.
- Page 65 There is no routine maintenance required for the Moxi V . In addition, there are no user serviceable parts with the exception of the ability to swap out the tray insert (see instructions above). Instrument repairs must be carried out by authorized personnel only. Page 61 ™...
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Page 66: Specifications For The Moxi V ™ And Cassettes
™ Specifications for the Moxi V and Cassettes System Performance Detection Parameters 3 (561nm/LP fluorescence, cell count and cell size) Excitation Laser Wavelength 532 nm Fluorescence Detection 561nm/LP filtered PIN diode Particle Count/Size Detection Impedimetric (Coulter Principle) Display 480 x 800 pixels, color touchscreen Resolution 1000 histogram bins Data Storage... -
Page 67: Ordering Information
Benchtop Dimensions (HxWxD) 5.8 in x 9.3 in x 8.7 in / 14.8 cm x 22.1 cm x 23.6 cm Weight 8 lbs / 3.6 kg Electrical Specifications Internal Battery Rechargeable 3.7V, 7500 mAh lithium ion Input: 100-240 VAC (50/60 Hz), 0.2 A AC Power Adapters (US, EU, and UK types) Output: 5 VDC, 1 A... -
Page 68: Technical And Customer Service
Moxi V USB Cable and Adapter MXA120 Cassette Tray Insert MXA090 Technical and Customer Service For technical service, contact ORFLO Technologies at 855-879-6694, Option 1 or email us at tech_support@orflo.com. For customer service, including sales, pricing, and order information, contact Gemini Bio Customer service at 800-543-6464 or customerservice@gembio.com. -
Page 69: Applicable Patents
What To Do If You Think You Have a Warranty Issue Please contact ORFLO promptly at 855-TRY-MOXI (855-879-6694, Option 1) or send an email to tech_support@orflo.com to describe the issue you are experiencing. Please be prepared to supply your name, contact information, the serial number of your product, and the original purchase date of the unit. - Page 70 A subsidiary of Page 66 ™ Moxi V User Guide...
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