FX500 Exchangeable Fluidics Cell Sorter

Software

A ribbon interface logically organizes functions to make them rapidly accessible. An experiment-centric approach makes the software easy to teach, learn, and use. The software is intuitive and supports actions from automated setup to acquisition and analysis. The software generates FCS 3.0 and FCS 3.1 files that also can be exported to third party analysis tools.

Software wizards include step-wise workflows that guide users through start up, multicolor compensation, sorting and system shutdown.


System Start Up

Upon start-up, the system initiation includes diagnostics that ensure all subsystems are working properly. Once verified, system status and a green ready message are displayed on the LCD monitor on the front of the instrument. The LCD monitor displays status information during operation.

fx500-automation-system-startup

The LCD monitor on the front of the FX500 displays status information during operation.


Alignment and Calibration

The system automatically and precisely aligns the sorting chip to the lasers and calibrates the droplets, side streams, and drop delay using AutoSetup beads.

software-calibration-setup-beads-package


Creating Experiments

Users can create an experiment by using templates from a selection of recent experiments. When a new template is created, guided dialogs prompt the user to select experiment settings which contains sample groups, tubes, and pulse parameters for data acquisition. Prior to starting acquisition, a user can set up compensation tubes.

fx500-software-creating-experiment

A template or a recent experiment on the left of the window displays the experiment structure on the right.


Data Analysis and Display

Data is displayed as dot plots and histograms on worksheets, and populations can be marked using gates. The software has a number of tools to select, adjust, label, and measure statistics of target populations. Data can be analyzed within the software or exported as FCS 3.0 or 3.1 formats to use with third party analysis software.

fx500-software-data-analysis-screen

The Experiments tab provides easy access to all lists of previous experiments as well as graphic and statistical tools for analysis.


Sorting

Once events are gated on plots, targeted cells can be sorted for further analysis. The software offers different sort modes to provide different levels of purity and yield as well as a single cell sorting mode to support optimal results. All sort controls are managed within a simple dialog.


Sort Deposition System and Index Sorting Software

The FX500 supports sorting into tubes by default. The Sort Deposition System enables sorting and precise deposition into 96- and 384-well plates and other plate formats. Index sorting software records the X and Y coordinates of each event sorted into a multi-well device. This feature allows researchers to track the scatter and fluorescence intensity of individual cells sorted into each well.

fx500-software-index-sorting


High-Purity Sorting of T Cells

In this data, cells from a buffy coat preparation from human blood were used at a nal concentration of 20 million/mL. Cells were stained with 10 microliters of human anti-CD45 APC (Cat # 2120060), anti-CD3 PE (Cat # 2102040), anti-CD8 FITC (Cat #2323520), and anti-CD4 PECy™7 (Cat #2102560) from Sony Biotechnology Inc. and were incubated for 30 minutes. The stained sample was run on the FX500 cell sorter using the 70µm sorting chip. At an event rate of 20,000 events/s, CD8+ and CD4+ cells were sorted with a purity of >99% and e ciency of >75%.

fx500-software-index-sorting