CellSimple™ Performance Beads were analyzed using the Open Flow Cytometry Application detecting in both green (525/45nm, left panel) and red (561nm LP, right panel) channels. Non-fluorescent and fluorescent beads were gated according to Median Fluorescence Intensity (MFI) and diameter (μm). Results are shown in the black rectangular boxes below the dot plots. Instrument screen shots are shown.
Prior to use, allow the beads to equilibrate to room temperature for at least 15 minutes. From the CellSimple™ Cell Analyzer main screen, choose the Open Flow Cytometry application and select both detection channels PMT1 (525/45 nm) and PMT2 (561 nm LP). The fluorescent gain setting should be set to “default”. Immediately prior to loading the Performance Beads, vortex them on high for 15 seconds then pipette 75 μl into the cassette and analyze. Do not dilute the aqueous solution. Gate the bead populations according to MFI and diameter (μm). Please see the CellSimple™ User Guide for more details about using the Open Flow Cytometry Application.
Store at 4° C and protect from light. Do not freeze as this will compromise bead performance. The Performance Beads are stable for at least 12 months when stored at the recommended conditions.
CellSimple™ Performance Beads are available as a quality control reagent for use with the CellSimple™ Cell Analyzer. The reagent contains a mixture of four beads: smaller red and larger green fluorescent beads, each with size-matched non-fluorescent beads. The four-bead mixture is run simultaneously and gated to determine fluorescence intensity, diameter and total count. The Performance Beads can be used on a regular basis to check instrument performance and reproducibility. Example data are shown in the attached figures.
The CellSimple™ Cell Analyzer is a benchtop instrument that utilizes a disposable thin-film cassette and a combination of a 488 nm laser, two photomultiplier tubes (525/45 nm and 561 nm LP filters), Coulter Principle-based cell measurements, and on-board software to provide easy-to-run applications and data analysis. Data acquisition occurs within approximately 10 seconds per test. The instrument relies on disposable cassettes for sample handling, which alleviates the need for flow cell cleaning and fluidics maintenance and the instrument is small enough to be portable between the lab bench and the hood. Applications include quantitative assessments of cell viability, apoptosis, other labeled antibody markers and single and multiplexed bead-based assays for protein and cellular analysis.
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