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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).

BD FACS Symphony A3 Configuration and Fluorochromes
Excitation LaserLongpass FilterBandpass FilterExamples of FluorochromesDetector
Blue (488 nm)505LP515/20BB515, FITC, AF 488, CFSE, GFP, TO-PRO-1BB515
600LP610/20BB630BB630
690LP710/50BB700, PerCP, PerCP-Cy5.5, PerCP-eFluor710, 7-AAD, PIBB700
770LP780/60BB790BB790
Red (637 nm)650LP670/30APC, AF633, AF647, TO-PRO-3APC
690LP710/50AF700, APC-Cy5.5, Vybrant® DyeCycle™ Ruby StainAF700
750LP780/60APC-CY7, APC-H7, APC-eFluor780APC Cy7
Yellow-green (561 nm)570LP586/15PE, AF555, AF546, dTomato, DsRed, RFPPE
600LP610/20PI, PE/Dazzle 594, PE-CF594, PE-Texas Red, mCherryPE
CF594
650LP670/30PE-Cy5, 7AAD, mPlumPE Cy5
750LP780/60PE-Cy7PE Cy7
Violet (405 nm)410LP431/28BV421BV421
410LP450/50BV421, DAPI, V450, Pacific BlueOptional
450LP470/15BV480BV480
600LP610/20BV605, Pacific Orange, eFluor605NCBV605
635LP670/30BV650BV
690LP710/50BV711, Yellow Dead CellBV711
750LP780/60BV786BV786
UV (355 nm)none379/28BUV396BUV396
410LP450/50DAPIDAPI
450LP515/30BUV496BUV496
550LP586/15BUV563BUV563
635LP670/30BUV661BUV661
690LP740/35BUV737BUV737
770LP820/60BUV805BUV805