Flow Cytometer

Fluorescence Sensitivity

Estimated detection limit is 750 molecules of equivalent soluble fluorescein

Fluorescence Resolution

Coefficient of variation in FL2-Area of <3%, full peak for propidium iodide-stained chicken erythrocyte nuclei

Forward and Side Scatter Sensitivity

Sensitivity enables the separation of fixed platelets from noise

Forward and Side Scatter Resolution

Scatter performance is optimized for resolving lymphocytes, monocytes, and granulocytes

Optical Platform

Fixed optical assembly

Lasers

15 milliwatt 488 nm, air-cooled Argon-ion laser; life expectancy >5,000 hours Optional second laser: nominally 635 nm

Beam Geometry

Prismatic expander and achromatic spherical lens provide 22 µm x 66 µm elliptical beam for argon ion laser, and nominally 15 µm x 61 µm elliptical beam for red diode laser

Optical Coupling

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

Background Rejection

Obscuration blade and slit minimize unwanted laser radiation at the detector

Forward Scatter Detector and Filter

High performance solid state silicon detector with 488 nm band pass filter for clear signal detection, and red diode laser signal rejection

Side Scatter Detector

High performance photomultiplier using Brewster angle beam splitter in the emission optical train

Fluorescence Detectors and Filters

Four high performance, high dynamic range photomultipliers with band pass filters: 530 nm (FITC), 585 nm (PE/PI), 661 nm (APC), and >650 nm (PerCP) with base unit, >670 nm (PerCP) with FL4 option

Fluorescence Sensitivity

Estimated detection limit is 750 molecules of equivalent soluble fluorescein

Fluorescence Resolution

Coefficient of variation in FL2-Area of <3%, full peak for propidium iodide-stained chicken erythrocyte nuclei

Forward and Side Scatter Sensitivity

Sensitivity enables the separation of fixed platelets from noise

Forward and Side Scatter Resolution

Scatter performance is optimized for resolving lymphocytes, monocytes, and granulocytes

Optical Platform

Fixed optical assembly

Lasers

15 milliwatt 488 nm, air-cooled Argon-ion laser; life expectancy >5,000 hours Optional second laser: nominally 635 nm

Beam Geometry

Prismatic expander and achromatic spherical lens provide 22 µm x 66 µm elliptical beam for argon ion laser, and nominally 15 µm x 61 µm elliptical beam for red diode laser

Optical Coupling

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

Background Rejection

Obscuration blade and slit minimize unwanted laser radiation at the detector

Forward Scatter Detector and Filter

High performance solid state silicon detector with 488 nm band pass filter for clear signal detection, and red diode laser signal rejection

Side Scatter Detector

High performance photomultiplier using Brewster angle beam splitter in the emission optical train

Fluorescence Detectors and Filters

Four high performance, high dynamic range photomultipliers with band pass filters: 530 nm (FITC), 585 nm (PE/PI), 661 nm (APC), and >650 nm (PerCP) with base unit, >670 nm (PerCP) with FL4 option

Cytometer

Dimensions (W x H x D)

91 x 64 x 61 cm (35.7 x 25.2 x 24 in)

Workspace Dimensions

Height (with flow cell access door open): 85 cm (33.5 in)

Power Requirements

100/115/230 VAC (50-60 Hz)

Fluorescence Threshold Sensitivities

FITC < 100 MESF

PE < 50 MESF

Forward and Side Scatter Sensitivities

Platelets can be resolved from noise

Forward Scatter Sensitivity

1 micron

Side Scatter Sensitivity

0.5 micron

Data Acquisition Channels

8 to 10 parameters: 6 to 8 fluorescent and 2 scatter parameters

Fluidics Cart

Dimensions (W x H x L)

79 x 64 x 61 cm (31.1 x 25.2 x 24 in)

BD FACSFlow Cubitainer Capacity

20 L

BD FACSClean Solution Cubitainer Capacity

5 L

BD FACS Shutdown Solution Cubitainer Capacity

5 L

Waste Tank Capacity

10 L

Performance

Fluorescence Sensitivity

FITC-A: fewer than 80 molecules of equivalent soluble fluorochrome (MESF-FITC) PE-A: fewer than 30 molecules of equivalent soluble fluorochrome (MESF-PE) Measurements performed using SPHERO™ Rainbow Calibration Particles RCP-30-5A

Fluorescence Resolution

Coefficient of variation PI: Area of <3%, full G0/G1 peak for propidium iodide (PI)-stained chicken erythrocyte nuclei (CEN).

Fluorescence Linearity

Doublet/singlet ratio of 1.95–2.05 for CEN stained with PI and excited with the 488-nm blue laser

Forward and Side Scatter Sensitivity

Enables separation of fixed platelets from noise.

Forward and Side Scatter Resolution

Scatter performance is optimized for resolving lymphocytes, monocytes, and granulocytes.

Data Acquisition Rate

Theoretical maximum electronic data collection of 70,000 events/second. Typical operation at 20,000 events/ second which requires a sample concentration of 2 x 107 cells/mL at an instrument flow rate of 60 μL/min.