Beam height: 9 ±3 μm Beam width: 65 ±7 μm
Power Out of the Laser Head
375 nm: >7 mW, top hat shape 405 nm: >50 mW, elliptical shape 445 nm: >20 mW, elliptical shape 488 nm: >20 mW, elliptical shape 561 nm: >50 mW, elliptical shape 633 nm: >18 mW, elliptical shape
Emission Optics Optical Coupling 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 and 462/52 band- pass filter for the 445-nm laser.
Side Scatter Detector
Photomultipler with a 488/10 bandpass filter for the 488-nm laser and a 445/15 bandpass filter for the 445-nm laser.
Fluorescence Detectors and Filters
Five fixed-fiber apertures for the 488, 633, 561, 445, and 405-nm lasers
The 445-nm laser focuses on the flow cell at the same location as the 488-nm laser. The 375-nm laser is air launched and focuses on the flow cell at the same location as the 405-nm laser. An octagon technology detector array enables user- defined detection configurations.
Wavelengths detected from 488-nm laser without 561-nm laser installed • 530/30-nm FITC • 585/42-nm PE
• 616/23-nm PE-Texas Red® • 695/40-nm PerCP-CyTM5.5 or PI or
675/20-nm PerCP-Cy7 • 760/60-nm PE-CyTM7
Wavelengths detected from 488-nm laser when 561-nm laser installed • 530/30-nm FITC • 695/40-nm PerCP-Cy5.5 or PI or
Wavelengths detected from 633-nm laser
• 660/20-nm APC • 780/60-nm APC-Cy7 or APC-H7
Wavelengths detected from 405-nm laser
• 450/50-nm BD HorizonTM V450, Pacific BlueTM, DAPI
• 510/50-nm BD Horizon V500, AmCyan
Wavelengths detected from 561-nm laser
• 582/15-nm PE • 610/20-nm PE-Texas Red®, Living
Colors®, mCherry, propidium iodide • 670/14-nm PE-CyTM5 or 710/50-nm
PE-Cy5.5 • 780/60-nm PE-Cy7 Wavelength detected from 445-nm laser • 510/80-nm, cyan fluorescent protein
Wavelengths detected from 375-nm laser
• 450/20-nm Hoechst Blue, DAPI • 670 LP Hoechst Red, PI
Filters and mirrors are user changeable. Additional detectors up to a total of 18 wavelengths can be added to the arrays.
488, 633, 561, 445, and 405-nm laser: Fiber optics steer the fixed alignment laser beams onto the expansion prisms to focus them on the cuvette flow cell.
Near UV (375-nm) laser: Air launched and focused on the cuvette flow cell
Sort Performance Drop Drive Frequency Range 1–100,000 Hz
Purity and Yield
At 70 psi and 90 kHz with an average threshold rate of 25,000 events per second, a four-way sort achieved a purity of >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 FACSTM 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
Loading 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)