Introducing the Worlds First “Smart Flow Cytometer?”

“Plug and Play” Simplicity

Moxi Flow is the World’s first SMART FLOW CYTOMETER. Moxi Flow is a fully automated, cassette-based flow cytometer that combines unparalleled ease of use with the precision and accuracy normally only associated with more expensive flow cytometers. This ultra-small instrument uses patented microfluidic thin-film cassettes that enable automatic load and measure operation. The Moxi Flow is also the only flow cytometer that auto-aligns the laser to each cassette to enable highly repeatable and robust results every time. No PMT gain adjustment is required. No warm up time required. Just insert a cassette, pipette your sample, and read the results in 10 seconds.

Versatile

Select from one of our standard applications (i.e., Cell Counts, Viability, Apoptosis) or run the system in open “2 Parameter Flow Cytometry” mode for the ultimate in assay flexibility. This miniature open platform can be used for your most common cell assay needs, or to develop sophisticated, custom assays right at your fingertips.

Access your FCS compatible Data with “USB-on-the-Go”

Moxi Flow generates data using the industry standard FCS 3.1 format so you can open and analyze your results using your ORFLO’s VESTIGO software (coming soon), or any other standardized flow cytometry analysis software. Data is transferred simply and easily to a PC or MAC using a mini-USB cable and the on board “USB-on-the-Go” functionality. No additional software is required to perform the data transfer.

Incredible Performance in a Small Package

? More accurate and repeatable than any other automated cell counter
? Cell Count with >95% accuracy
? Viability with PI
? Apoptosis with Annexin V
? Precise cell/particle (Coulter Principle)
? Blood analysis (RBC counts, WBC counts, CD4 counts, etc.)
? Somatic Cell Counts in Milk

Perform state of the art flow cytometry with combined fluorescence and impedance detection with Orflo’s Moxi Flow platform. 

Applications  
Moxi Flow comes with pre-loaded applications that are optimized for each assay. New applications are always under development and will be released, free of charge, to existing Moxi Flow owners via downloadable OS updates. The current applications are:

? Viability with PI ( MoxiCyte Viability Kit Protocol)
? Apoptosis ( Moxi Flow - Annexin V-PE Staining Protocol)
? Open Flow Cytometry ( Moxi Flow - CD4-PMBC Staining Protocol)
? Size Histogram
? Fluorescent Bead Assay

Viability Count (PI) -  Obtain Cell Counts with Viability in 95% accuracy).
Perform routine cell counts with viability with a never before combination of simplicity, accuracy, and thoroughness. Simply select the “Viability Count (PI)” app, insert a cassette, pipette your cells stained with Propidium Iodide, and let the Moxi Flow take it from there. Better than 95% accurate data is generated automatically in both a color density dot plot and ORFLO’s unique Color Viability Histogram? that displays both the live cells (green) and dead cell (red) on one, easy to read size histogram. Because the Moxi Flow measures cell volume using the Coulter principle, actual cell size is measured, not just an estimate from forward or side scatter.

 MoxiCyte Viability Kit Protocol

Fluorescence vs. size scatter plot of a partially-killed (60C, 30min), propidium iodide (PI) - labeled 3T3 population.

Size histogram of a partially-killed (60C, 30min), propidium iodide (PI) - labeled 3T3 population with nonviable cells in red and viable cells in green.

Fluorescence vs. size scatter plot of a Camptothecin-treated (50uM, ~4hr), propidium iodide (PI) - labeled CHO-K1 population.

Size histogram of a Camptothecin-treated (50uM, ~4hr), propidium iodide (PI) - labeled CHO-K1 population with non-viable cells in red and viable cells in green.

Fluorescence vs. size scatter plot of a partially-killed (60C, 30min), propidium iodide (PI) - labeled Jurkat population.

Size histogram of a partially-killed (60C, 30min), propidium iodide (PI) – labeled Jurkat population with non-viable cells in red and viable cells in green.

Fluorescence vs. size scatter plot of Ficoll-Paque purified, propidium iodide (PI) - labeled PBMC population.

Size histogram of Ficoll-Paque purified, propidium iodide (PI) – labeled PBMC population with non-viable cells in red and viable cells in green.

Apoptosis -  Study more advanced mechanism of cell death using the Apoptosis app. ORFLO has optimized system performance to run with our Apoptosis Assay Kit that utilizes Biolegend’s Annexin V-PE conjugated dye. Simply prepare your cells as instructed in the Apoptosis Assay Kit, select the Apoptosis app from the Moxi Flow home page and you are automatically running sophisticated apoptosis flow cytometry assays. The Color Density Scatter Plots are easily gated on the unit and percent apoptotic cells is automatically calculated.

 Moxi Flow - Annexin V-PE Staining Protocol

Fluorescence vs. size scatter plot for Annexin-PE labeled CHO-K1 cells following ~4hr treatment with 50uM Camptothecin.

Size histogram Annexin V-PE+ (red) and Annexin V-PE- (green) CHO-K1 cells following ~4hr treatment with 50uM Camptothecin.

Kinetic progression of apoptosis in Camptothecin-treated (50uM) Jurkat cells. Images show Fluorescence vs. size scatter plots at post-treatment times of: 2 hour, 6 hour, and 7 hour.

Size Histogram -  Run your Moxi Flow like a Moxi Z. The Size Histogram app runs the system using the electric impedance detection mode only. Volumetric cell counts with size discrimination are automatically generated and displayed in histogram format. Particles down to 3μm and up to 25μm (effective diameter) can be analyzed depending on the cassette selected.

Size histogram of a Camptothecintreated (50uM,~24hr) Jurkat E6.1 cell culture illustrating the size shift commonly exhibited by cells undergoing apoptosis (left peak) vs the non-apoptotic (right, redcurve fit peak) cells.

Size histogram of white blood cell (WBC) nuclei from Zap-oglobin II treated (4 drops), diluted (40uL Blood:15mL Orflo Diluent) whole blood. This technique can be used to provide rapid total WBC counts.

Size histogram of precision – sized (NIST) 4.17uM polystyrene beads.

Open Flow Cytometry -  Design your own flow cytometry assay by selecting flourophores that are excited at 532nm. When this app is selected, researchers have the option of running the Moxi Flow at three different sensitivity levels (Low, Default, and High) to enable the system to match the sensitivity of the fluorophore being used. Because actual cell volume is measured simultaneously with the labelled flourescent activity (instead of the traditional estimate of cell volume, i.e., forward scatter), Moxi Flow will enable an array of experiments to be conducted that have, to date, not been possible.

 Moxi Flow - CD4-PMBC Staining Protocol

Fluoresence vs. size scatter plot of anti-human CD4 – PE labeled PBMC’s (Ficoll-purified).

Size histogram of anti-human CD4 – PE labeled PBMC’s (Ficoll-purified) with CD4+ cells in red and CD4- cells in green. Left-most cell peak is RBC- contaminated population, middle peak is predominantly lymphocyte cells and right peak is predominantly monocyte cells.

Fluoresence vs. size scatter plot of anti-human CD4 – PE labeled PBMC’s (Ficoll-purified).

Fluorescent Bead Assay-  The Flourescent Bead app lets researchers run the Moxi Flow in a mode optimized for Fluorescent microspheres. ORFLO will soon be announcing the release of our proprietary Moxi-Plex? kits and corresponding user interface to allow users to perform sophisticated, quantitative, multi-plex protein assays right on the unit. The Moxi-Plex update will be offered free of charge and is scheduled to be released in 2014.

Example fluorescence vs. size scatter plot of an eleven bead mix of varying sizes (3.8um, 5.05um, 7.8um, 10.6um, and 10.8um) and intensities designed to illustrate the Multiplexing capabilities of the Moxi Z.

Fluorescence vs. size scatter plot of Spherotech ultra-rainbow bead (URB) fluorescence calibration beads. Five separate bead intensities (no blank) are present in this mix and clearly resolved by the Moxi Flow.

Fluorescence vs. size scatter plot with Fluoresence histogram (blue) and size histogram (red) overlay of BD Quantibrite calibration beads. Beads represent MESF values (in order of increasing intensity) of 474, 5359, 23843, and 62336 molecular equivalent soluble fluorophore (MESF) values.

Future Apps -  In addition to Moxi-Plex, ORFLO is actively developing applications, and corresponding reagent kits, that will be released to Moxi Flow owners free of charge. Applications currently under development include, CD4+ T cell counts, cell cycle, and Micobe- Check (an app that identifies undesired microbes in cell cultures). Please check our website for the latest Moxi-Flow firmware updates containing new applications.

When you use the new Flow Cassettes, you will enjoy the benefit of having a brand new Flow Cell, perfectly aligned, for every experiment

Higher Performance & Simplicity in One Disposable Cassette.

The unique, patent-protected thin-film Flow cassettes provide highly accurate, repeatable cell counts, cell size, and one color fluorescence analysis in 5 to 15 seconds (depending on cassette type). Inspired by the gold-standard Coulter Principle used in high-end counting systems worldwide, the Flow cassettes move thousands of cells per test through a cell sensing zone to accurately capture the impedance-based volumetric measurement of each inpidual cell in the sample, while simultaneously measuring each cell fluorescently. Flow cytometry methods for cell counting of inpidual cells is proven to be over 95% accurate versus a 75% accuracy with image-based cell counting methods.

No system contamination and integrated pre-filter for clogging prevention.

Flow cassettes are designed with a unique “cell sieve” to minimize cell clumping and clogging. In addition, Flow cassettes contain 100% of your sample within the cassette body, eliminating the possibility for system contamination and the corresponding need for sterilization. Cassettes are simply and easily discarded when testing is finished.

Used cassette indicators.

Each Flow cassette offers two inpidual tests that can be used independently. Moxi Z and Moxi Flow systems will also alert the user if a used cassette is mistakenly inserted in the unit.

Two cassette types for maximum flexibility and performance

For optimum performance for smaller calls and microbeads, we recommend using the MF-S cassettes. For larger cells and more adherent cell lines, we recommend using the MF-M cassettes. 
See www.orflo/moxiz/applications for a list of cell types and corresponding cassette types.

How it Works.

The operating principle behind the Moxi Flow Smart Flow Cytometer is a unique combination of Coulter-style cell size determination with simultaneous fluorescence detection. As cells flow single file through the microfabricated single-use flow cell the volume of each particle is measured at the exact same time as their primary fluorescence is measured using a 532nm solid state diode laser. Thousands of cells are measured in the 10 second read time and the data are plotted in a gradient density scatter plot as Cell size (volume) vs. Fluorescence (PMT voltage). Gating is easily performed on the unit using a interactive touch display, and the resulting live/dead ratios are automatically calculated (depending on the app selected). The analyzed data can also be displayed as a two color size histogram. Total volumetric cell counts are automatically determined for each test by precisely measuring the volume of fluid being analyzed.

Operation

Simple 3 step operation.

Step 1: Select desired app, insert the cassette and close the doors.

Step 2: Once auto-alignment is complete, open the top door and pipette 50μl of labelled sample into the cassette.

Step 3: Close the top door, assays run automatically and results are generated in 10 seconds.

Note: Each cassette holds 2 tests. When Sample 1 is complete, simply re-insert other end of cassette into Moxi Flow, and load Sample 2.

Data

Data can be displayed on the unit in both a color density scatter plot and a two color size histogram. Simply drag gates using the intuitive touch display for instant live/dead ratio calculations and each of the gated volumetric cell counts (i.e., total population, live population, and dead population (Viability App). The mean cell volume for the gated populations is also automatically displayed on the unit.

Results from each test are stored in the standard FCS 3:1 format and can be viewed using any flow cytometry analysis package. The actual Moxi Flow screenshots from each assay (dot plots and histograms) are also stored in bitmap format for use online. Hundreds of files can be stored on each Moxi flow and are easily transferred to a Mac or PC using USB on-the-go. No aditional software is required.