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

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. Shay Kirzner before ordering this system.

 

 

Related Documents

AriaIII configuration