BD FACSymphony A3 Cell Analyzer

The BD FACSymphony™A3 Cell Analyzer is a novel high parameter system that offers qualitative and quantitative analysis of cells and other particles using multi-parametric analysis. This instrument is configured for five laser (UV 355 nm, Violet 405 nm, Blue 488 nm, Yellow-Green 561 nm, Red 637 nm) analysis and can simultaneously acquire and distinguish between 23 different fluorescent signals in addition to the forward and side scatter parameters allowing for a 25-parameter definition of the cell. The FACSymphony has the capacity to acquire up to 40,000 events per second allowing for a rapid acquisition of samples.  Additionally, the BD High-Throughput Sampler (HTS) option can be used to acquire samples from a 96 or 384-well plate. The FACSymphony is operated using FACSDiva 9.1 software.  FCS file versions 2.0, 3.0, and 3.1 can be exported for data analysis.

Sample Preparation for Flow Cytometry Analysis (FACSymphony A3) 
 
For flow cytometry analysis, the minimal volume of sample required is 0.5 mL; the optimal concentration is between 0.5-1 x 106 cells per sample. Bring the resuspended cells in a 5 ml 12 x 75mm polystyrene round-bottom tubes (BD Falcon cat. number 352054). Samples must be in a single cell suspension. If they are not, you run the risk of clogging the instrument since the tubing and nozzle through which the cells pass is very narrow. 
 
If you see clumps or aggregates of cells and/or debris in your sample, it will not run properly. Adding 1mM EDTA should prevent the formation of cell clumps. In order to remove clumps or aggregates, the sample should be filtered prior to running. We recommend using Falcon 5 ml polystyrene round-bottom tubes with cell strainer cap (BD Falcon cat. number 352235).