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  Institute of Cell and Molecular Science
Flow Cytometry Core Facility equipment, images used with permission by BD
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BD FACSAria IIIu sorter
Specification Comments
Performance
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 20,000 events/sec
Excitation optics
Optical platform Fixed optical assembly, upon cuvette flow cell
Lasers

20mW Coherent Sapphire blue 488nm: 530/30nm: 695/40nm
30mW Yellow-Green laser: 575/26nm; 610/20nm; 695/40nm; 780/60nm.
25 mW 405 Coherent VioFlame: 440/40nm; 530/25nm; 610/20nm; 670/20mn; 695/40nm; 780/60nm.
20 mW 635 JDS Uniphase HeNe; 660/20nm; 695/40nm; 780/60nm.

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 17 parameters, 15 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 Time available correlated to any parameter
Channel triggering Available on any laser signal
Data management
Central processing Dual Intel Core 2 CPU
Data storage 2GB RAM, 300-GB internal hard drive, 3.5-inch floppy drive, CD-RW drive
Data File Structure Flow Cytometry Standard 3.0 or 2.0
Software BD FACSDiva software v6.1.03
Fluidics
General Fluidics cart   provides sheath and cleaning fluid to instrument and receives waste
No air or vacuum required
Adjustable sheath pressure
Sample flow rate Adjustable
Sample injection chamber
Sample input agitation Adjustable through software
Temperature control Software adjustable, 4, 20, 37 and 42 o C
Aerosol management
Standard feature
Aerosol Management System - AMS- fitted with Buffalo HEPA filter

Sample injection chamber and sample collection tube area provide a sealed area to minimize aerosols.
Sort chamber is under negative pressure as per Class II Laminar Flow Cabinet (tested) from the Aerosol Management System (AMS) to remove aerosols generated to a HEPA filter.

Sort performance
Sort collection devices

Two way sorting and four way sorting in to sample tubes

70um, 85um and 100um 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

 
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by Gary Warnes. © Queen Mary, University of London 2007
Institute of Cell and Molecular Science, The Blizard Building, Barts and The London School of Medicine and Dentistry, 4 Newark Street, London E1 2AT, UK Tel: +44 (0)20 7882 2483, Fax: +44 (0)20 7882 2200