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BD FACS Aria III 3 lasers (5B, 2R, 2V) with ACDU
BD Workstation w/ FACSDiva 6.1.3 and Dongle
Fixed optical alignment of all Class IIIb lasers upon the cuvette flow cell. The 488- nm and 633-nm lasers come standard. All other laser choices are optional. All are solid state except the 633 nm, which is gas.
Beam height: 9 ±3 µm
Beam width: 65 ±7 µm
Power out of the laser head
355 nm: >15 mW, elliptical shape
405 nm: >50 mW, elliptical shape
488 nm: >20 mW, elliptical shape
561 nm: >50 mW, elliptical shape
633 nm: >18 mW, elliptical shape
488, 633, 561 and 405-nm laser: Fiber optics steer the fixed alignment laser beams onto the expansion prisms to focus them on the cuvette flow cell.
UV (355-nm) laser: Air-launched and focused on the cuvette flow cell at the same location as the 405-nm lasers.
The quartz cuvette flow cell is gel-coupled by refractive index-matching optical gel to the fluorescent objective lens for optimal collection efficiency.
Numerical aperture: 1.2
Forward scatter detector and filters
Photodiode with 488/10 bandpass filter for the 488-nm laser.
Side scatter detector
Photomultiplier with a 488/10 bandpass filter for the 488-nm laser.
Fluorescence detectors and filters
Four fixed-fiber apertures. The 488-nm, 633-nm and 561-nm lasers have their own aperture and fiber. The 405-nm and 355-nm lasers share an aperture and fiber.
An octagon technology detector array enables user-defined detection configurations.
Filters and mirrors are user-changeable. Additional detectors up to a total of 18 wavelengths can be added to the arrays.
Wavelengths detected from 488-nm laser without 561-nm laser installed
616/23-nm PE-Texas Red®
695/40-nm PerCP-Cy™5.5 or PI or
Wavelengths detected from 488-nm laser with 561-nm laser installed
695/40-nm PerCP-Cy5.5 or PI or 675/20-nm PerCP
Wavelengths detected from 633-nm laser
780/60-nm APC-Cy7 or APC-H7
Wavelengths detected from 405-nm laser
450/50-nm BD Horizon™ V450, Pacific Blue™, DAPI
510/50-nm BD Horizon V500, AmCyan
Wavelengths detected from 561-nm laser
610/20-nm PE-Texas Red®, Living Colors®, mCherry, propidium iodide
670/14-nm PE-Cy™5 or 710/50-nm PECy5.5
Wavelengths detected from 355-nm laser
450/20-nm Hoechst Blue, DAPI
670 LP Hoechst Red, PI
The fluidics cart provides sheath and cleaning fluids to the instrument and receives waste. No air or vacuum required. Room air can be used if desired. Sheath pressure is adjustable from 5 to 75 psi.
Autoclavable 10-L sheath and waste containers and 5-L cleaning reservoirs provided.
Sample flow rates
Adjustable dynamic range of sample flow rates.
Fluidic cleaning modes included (software)
Automated startup and shutdown
Clean flow cell
Prepare for aseptic sort
70-, 85-, 100- and 130-µm, removable and can be sonicated.
A registered key-fit position at the bottom of the cuvette provides fixed stream alignment.
Sample collection cooling/heating
Refrigerator/heater option is available to provide cooling or heating for sort collection into tube holders, multiwell plates and slides.
Operating temperature range: -20°C to 120°C
Automated cell deposition unit (ACDU)
ACDU for slide and plate sorting: 6-, 24-,48-, 96- and 384-well plates.
Measurements performed at 70 psi and 90 kHz using SPHERO™ Rainbow Calibration Particles (RCP-30-5A)
FITC: 85 molecules of equivalent soluble fluorochrome (MESF-FITC)
PE: 29 molecules of equivalent soluble fluorochrome (MESF-PE)
Coefficient of variation (CV)
PI: Area, <3.0%, full G0/G1peak for propidium iodide (PI)-stained chicken erythrocyte nuclei (CEN)
Hoechst: Area, 98% and a yield >80% of Poisson’s expected yield. Higher threshold rates up to 70,000 events per second can be achieved without affecting purity; however, yield will decrease based on Poisson statistics.
Sorts were performed at a variety of sheath pressures using cell lines and human peripheral blood mononuclear cells (PBMCs). All sorts resulted in cells that proved viable and proliferated for several days post-sort.
Sort collection devices
Two-way sorting: 12 x 75 mm, and 15 mL
Four-way sorting: 1.5 mL microtube and 12 x 75 mm
Sort collection cooling
Water recirculator for refrigeration/heating (optional)
BD FACS™ Accudrop
Red diode laser provided for fully automated drop-delay determination
Automated drop breakoff monitoring
Automated clog detection and sort tube protection system using Sweet Spot technology
10-MHz Analog-to-Digital converter.
Pulse sampling is precisely matched to particle flow rate in the cuvette. Particles travel slower compared to conventional stream-in-air sorters. This increases the light collected, resulting in better sensitivity. High-speed sorting is achieved by accelerating the stream through the nozzle, achieving drop rates comparable to stream-in-air sorters. The flow cell design and electronics are matched to maximize signal while maintaining maximum sort speed, purity and yield.
Data acquisition channels
20 parameters, 18 fluorescent and 2 scatter
No limit to inter- and intra-beam compensation
Height, area and width measurements available for any parameter. Ratio measurements are also available.
Time can be correlated to any parameter for kinetic experiments or other applications.
Available for any parameter from any lasers with the ability to use multiple thresholds from different lasers simultaneously.
Sample input sizes
Microtubes, 12 x 75 mm and 15 mL
Polystyrene or polypropylene tubes can be used.
Sample input agitation
Adjustable through the software to keep sample constantly suspended
Sample input, software-adjustable: 4, 20, 37 and 42°C
Sample output for sort collection: water recirculation unit (optional)
BD FACSAria III sensor (cell sorter):
28 H x 28 D x 48 W in. (71 x 71 x 122 cm)
400 lb (181.4 kg)
BD FACSAria III fluidics cart (wet cart):
26 H x 26 D x 32 W in. (66 x 66 x 81 cm)
180 lb (81.7 kg)
Temperature operating range
Between 17.5°C (63.5°F) and 27.5°C (81.5°F)
5100 BTU per hour
Operation at 100/115/230 VAC and 50 or 60 Hz
Maximum power: 1,500 watts
Water supply: None required
Air supply: None required
BD FACSAria III instrument and computer table: 33.5 H x 53 D x 105 W in. (85 x 135 x 267 cm) 100 lb (45.4 kg)
Aerosol Management Option (AMO)
The BD FACSAria III features an enclosed pathway from the sample injection chamber to the sort collection tubes. For an added level of aerosol management, the BD Aerosol Management Option (AMO) evacuates the sort collection chamber and traps aerosolized particles during sorting. It is equipped with a 0.01- μm size ultra-low penetrating air (ULPA) filter to trap aerosolized particles. Evacuates the volume of the ACDU chamber area 16 times per minute in normal evacuation mode and 69 times per minute in rapid evacuation mode.
When operated under normal and stressed conditions (mimicking a clog), <3 Glo Germ™ particles were identified outside the ACDU sort collection chamber. Glo Germ particles, developed by Glo Germ in Moab, Utah, have been shown to provide good visualization of aerosol deposition in normal and mock failure modes by Oberyszyn and Robertson (Cytometry. 43:217-222, 2001)