1. Optics
1) Excitation
Optics
- Fixed
optical alignment of all Class IIIb lasers upon the cuvette flow cel
- 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 ¡¾3 ¥ìm
- Beam width:
65 ¡¾7 ¥ìm
2) Emission
Optics
2-1) 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
2-2) Forward
Scatter Detector and Filters
- Photodiode with 488/10 bandpass
filter for the 488-nm laser and 462/52 bandpass filter for the 445-nm laser.
2-3) 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
2-4)
Fluorescence Detectors and Filters
- Five fixed-fiber apertures for the
488, 633, 561, 445, and 405-nm lasers
2. Fluidics
1) Nozzles
- 70, 85, 100¥ìm, removable and can be sonicated
2) Automatic Cell Deposition Unit
- ACDU for
slide and plate sorting : 6, 24, 48, 96, and 384-well plates
3. Performance
1) Sort
Performance
3-1) Drop Drive Frequency
- Range : 1 -
100,000 Hz
3-2) 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
3-3) Viability
- 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
3-4) 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
3-5) Sort Collection Cooling
- Water recirculator for refrigeration/heating |