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FACS Aria–IIIu Sorter
Description
BD FACSAria-IIIu cell sorter is a new digital sorter enabling high-speed sorting and multicolor analysis.
The instrument accommodates 4 air-cooled lasers at 488-nm, 633-nm, 407-nm and a new laser – 561 nm
Sorting at up to 15,000 events/second
Multicolor analysis of up to 15 parameters
Up to four-way bulk sorting devices for a variety of tube sizes.
Automated Cell Deposition Unit (ACDU) for sorting to multi-well plates or microscope slides
Features
Optical System: Patented BD OctagonTM and TrigonTM technology.
Laser excitation and filters (see FACSAria III configuration in the related links):
Lasers: 488-nm, 633-nm, 561nm, and 407-nm.
15 parameters: forward scatter, side scatter, 5 fluorescent detectors of 488-nm, 2 fluorescent detectors of 633-nm, 4 fluorescent detectors of 561-nm, and 2 fluorescent detectors of 407-nm.
Laser line/color (Excitation) |
Detector | Emission Filter (nm) | Example of available markers |
407 nm Violet
|
A | 695 /40 (Far red) |
QD-710 |
B | 450/40 (Blue) |
DAPI, Hoechst 33258, Pacific Blue, BV421, Alexa Fluor 405 |
|
488 nm Blue
|
A | 780 / 60 (Infra-Red) | PE-Cy7 |
B | 695 / 40 (Red-FarRed) |
PE-Cy5, PerCP, PerCP-Cy5.5, 7-AAD, PI |
|
C | 616/23 (Red) | PE-Texas Red, PI | |
D | 585 / 42 (Orange-Red) |
PE, PI |
|
E | 530 / 30 (Green) |
Alexa Fluor 488, FITC, GFP, CFSE, Sytox Green, YFP |
|
F | 488 / 10 | SSC | |
FSC | |||
561 nm Yellow Green
|
A | 780 / 60 (Infra-Red) | PE-Cy7 |
B | 670/14 (Far Red) |
7-AAD , PE-Cy5 |
|
C | 610/20 (Red) | mCherry, PE-CF594 , PI | |
D | 582/15 (Orange-Red) |
Cy3, PE |
|
633nm Red
|
A | 780/60 (Infra-Red) |
APC-Cy7 |
B | 660/20 (Red) |
APC |
Additional available filters:
Mirror | 450LP | 5005LP | 530LP | 635LP | 685LP | 735LP | ||
Band pass | 405/20 | 525/50 | 530/30 | 585/42 | 610/20 | 670/14 | 675/20 | 695/40 |
Specifications and performance
Fluorescence Sensitivity
FITC: 125 molecules of equivalent soluble fluorochrome (MESF-FITC)
PE: 125 molecules of equivalent soluble fluorochrome (MESF-PE)
Fluorescence Resolution
Coefficient of variation PI-Area of <3.0%, full G0/G1 peak for propidium iodide (PI)-stained chicken erythrocyte nuclei
Coefficient of variation Hoechst-Area of <3.5%, full G0/G1 peak for Hoechst-stained chicken erythrocyte nuclei
Fluorescence Linearity
Doublet/singlets ratio for CEN stained with PI = 1.95-2.05 detected off the 488-nm laser or Hoechst = 1.95-2.05 detected off the 407-nm laser
Forward and Side Scatter Sensitivity
Sensitivity enables separation of fixed platelets from noise, identification of bacteria, and 0.5-micron beads.
Forward and Side Scatter Resolution
Scatter performance is optimized for resolving lymphocytes, monocytes, and granulocytes.
Sample Acquisition Rate
Maximum acquisition rate: 70,000 events per second with 8 parameters, 12 compensation pairs. Maximum rate < 70,000 events per second with more parameters, > 100,000 events per second with fewer parameters.
Sorting options:
Two- and four-way sorting into microtubes, 12 X 75, and 15 mL tubes
ACDU (automated cell deposition unit) for sorting into multi-well plates and onto microscope slides
Sample injection chamber for various sample input tubes, including microtubes, 12 X 75, and 15 mL tubes. Includes sample agitation and temperature control.
Signal Processing
- Digital electronics with 10 MHz sampling frequency
- Up to sixteen acquisition channels with up to four spatially separated laser beams
- 18-bit data acquisition for area signals
- Inter- and intra-beam compensation on-line and post-acquisition off-line
- Height, width, and area measurement available for all parameters
- Time parameter available for kinetic studies (Calcium)
- Ratio measurements between any two parameters from any laser
Nozzle selection and design
An expanded set of nozzles lets users sort a wide range of particle sizes. Four nozzle sizes are supported: 70, 85, 100, and 130 microns.
Working policy
Users who wish to work on the FACS-Aria-IIIu must contact Dr. Aviv Lutaty before ordering this system.
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