FACS Aria–IIIu Sorter

Description

BD FACSAria-III 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

Digital acquisition rates of up to 70,000 events/second

Multicolor analysis of up to 15 parameters

Two- and four-way bulk sorting devices for a variety of tube sizes

Automated Cell Deposition Unit (ACDU) for sorting to multiwell plates or microscope slides

 

Features

488-nm, 633-nm, and 407-nm lasers

11 parameters, Forward scatter, side scatter, 5 fluorescent detectors off 488-nm, 2 fluorescent detectors off 633-nm, and 2 fluorescent detectors off 407-nm

(See more information in: BD FACS Aria-II configuration, at the related links)

  • Lasers and filters:

 

Laser line/color
(Excitation)
Detector Emission Filter (nm) Example of available markers

488 nm / Blue

Coherent Sapphire solid-state laser,

22 mW

 A  780 / 60 (Infra-Red)  PE-Cy7
 B   695 / 40 (Red-FarRed)

PerCP-Cy5-5

 C 610/20 (Red) PE-TxRed
 D   585 / 42 (Orange-Red)

PE, PI

  E 530 / 30 (Green)

Alexa Four 488, FITC, GFP,

CFSE, Sytox Green, YFP,

BOBIPY, Oregon Green,

Pico Green

F (SSC)  488 / 10 SSC
 FSC  FSC

633 nm / Red

JDS Uniphase Helium Neon laser, 20 mW

 A   780 / 60 (Infra Red)   APC-Cy7
 B 710/20 (Far-Red) APC-Cy5.5
 C  660 / 20 (Red)  APC, Alexa Flour 647

407 nm / Violet

Coherent VioFlame solid-state laser,

25 mW

 A   530 / 30 (Green)

AmCyan, Cascade Yellow,

Alexa Flour 430

 B   488/20

Cascade Blue

 C  450 / 40 (Blue)

Cascade Blue, Pacific Blue,

Hoechst, DAPI, Alexa Fluor 405

 

New 561 nm laser –

At the beginning of November 2013, the FACS Aria II was upgrade to FACS Aria III.  This upgrade includes the replacement and incorporation of a next-generation cuvette in the flow cell. Its design helps ensure that lasers are precisely focused on the sample stream, that they generate the greatest signal, and that the maximum amount of emitted light is collected. Furthermore, we added an additional laser, the 561nm (Yellow-Green) laser that provides the capability to efficiently excite fruit fluorescent proteins such as mCherry and mTomato as well as YFP.

Another advantage of the 561nm laser is the ease of separation of two commonly used fluorochromes, FITC and PE, resulting in the elimination of spillover between their emission spectra.

 

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 multiwell 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.

Digital Electronics

The BD FACSAria II cell sorter uses digital electronics to acquire data at rates up to 70,000 events per second for up to 8 parameters, with linear results over a broad dynamic range.

 

Applications

Aseptic sorting of events.

 

 

Working policy

users who wishes to work on the FACS-Aria-II must contact Dr. Shay Kirzner before ordering this system.

 

 

Releted Documents

 

Releted Links

BD FACS Aria-II