Immune Monitoring Laboratory at Dartmouth-Hitchcock Medical Center


CMS - 1.6.8 - Pouebo

FACSAria II cell sorter (336W Borwell)




Optical Bench

Before you decide on a fluorochrome for a flow cytometric assay, you need to know:
  • the wavelengths of the lasers in the cytometer
  • the wavelengths of the filters in front of the photomultiplier tubes (PMTs) in the cytometer
  • the absorption and fluorescence wavelengths of fluorochromes to be used

These can be simulated using the BD fluorescence spectrum viewer.


There are 2 possible filter configurations for the violet laser PMTs using either V2(i) or V2(ii) filters:

Blue laser configurartion:

Red laser configuation:

FACSAria optical bench:



Technical Specifications





Fluorescence sensitivity

FITC 125 molecules of equivalent soluble fluorochrome
PE 125 molecules of equivalent soluble fluorochrome

Fluorescence resolution

CV < 3% for PI labelled chick erythrocyte nuclei
CV < 3% for Hoechst labelled chick erythrocyte nuclei

Forward and side scatter sensitivity

Sensitivity enables the separation of fixed platelets from noise
Identification of bacteria and 0.5 micron beads

Forward and side scatter resolution

70,000 events/sec

Excitation optics

Optical platform

Fixed optical assembly, upon cuvette flow cell


20mW Coherent Sapphire 488nm, air cooled, argon ion laser
25 mW 405 Coherent VioFlame
20 mW 635 JDS Uniphase HeNe

Emission optics

Optical Coupling

Quartz cuvette is coupled to emission lens by refractive index-matching optical gel for optimum collection efficiency

Fluorescence Detectors

Three fixed fibre apertures

Twelve fluorescence detectors

High performance, high dynamic range photomultipliers with filters

Steering Optics

Fibre optics steer the three laser beams onto the beam expansion prisms, and then are focused upon the cuvette flow cell

Signal processing

Workstation resolution

Five decades for peak detection, offset less than 1/10,000 full scale

Data acquisition channels

13 parameters, 11 fluorescent and two scatter

Dynamic range

18-bit data acquisition
262,144 linear channels displayed in a 5-decade logarithmic display

Pulse processing

Width and area measurements available for all fluorescence parameters
Ratio measurements for intra-laser parameters


Time available correlated to any parameter

Channel triggering

Available on any laser signal

Data management

Central processing

Dual Intel Xeon processors

Data storage

Nine and 80-GB internal hard drive, internal drive with 250-MG removable disk storage space capacity, 3.5-inch floopy drive, DVD/CD-RW combo drive, Iomega Zip 750 MB

Data File Structure

Flow Cytometry Standard 3.0 or 2.0


BD FACSDiva software v3.2



Fluidics cart provides sheath and cleaning fluid to instrument and receives waste
No air or vacuum required
Adjustable sheath pressure

Sample flow rate


Sample injection chamber

Sample input agitation

Adjustable through software

Temperature control

Software adjustable, 4, 20, 37 and 42 o C

Aerosol management

Standard feature

Sample injection chamber and sample collection tube area provide a sealed area to minimise aerosols

Sort performance

Sort collection devices

Two way sorting and four way sorting in to sample tubes

70um, 100um and 130um diameter nozzle sizes available for sorting at 20,000, 10,000 and 5,000 cells per second at 70, 20 and 12 psi.

Automatic cell deposition unit allows for sorting onto slides or plates
Water re-circulator for refrigeration/heating collected sample - top