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BD Biosciences LSR II Flow Cytometer
The BD Biosciences LSR II is a multi-laser flow cytometer
that utilizes fixed-alignment lasers that transmit light reflected by mirrors
through a flow cell to user-defined octagon and trigon arrays. These
fluorescence signals are collected and translated to electronic signals then
converted to digital data. The BD LSR II utilizes innovative optics and digital
electronics to provide highly flexible yet powerful performance with a greater
sensitivity, which yields more information from each sample.
The BD LSR II can be configured with one to four
fixed-aligned, air-cooled lasers; this configuration features three lasers for
eleven color excitation. These solid state lasers consist of the Coherent Sapphire,
Coherent VioFlame Plus, and Lightwave Xcyte, all designed to consume less
power, produce less heat, and have a lower overall lifetime cost compared to
gas-filled lasers. The Coherent Sapphire (blue, 488 nm) produces no plasma
glow, lowering noise during excitation. The Coherent VioFlame Plus (violet, 405
nm) excites Cascade Blue, Pacific Blue, and Alexa Fluor 405, along with BD
Clontech Living Colors GFP, YFP, dsRed, and CFP. The Lightwave Xcyte (UV, 355 nm) allows
routine experiments to be performed on samples with UV dyes, such as DAPI,
Hoechst, and Indo-1.
The collection optics includes patented octagon and trigon
optical arrays, which increase sensitivity and flexibility for more sample
information. The 488-nm octagon array can collect up to seven colors along with
side scatter, while the 633-nm, 405-nm, and 355-nm trigon arrays can detect up
to three colors. Acting upon the principle that the reflectance of light is
more efficient than the transmittance, more colors can be detected with minimum
light loss.
The BD LSR II utilizes BD FacsDiva software, which provides a
means for experiment design, instrument control, and data analysis. FacsDiva
allows for improved workflow, featuring reusable acquisition templates, an
auto-spillover matrix for automated compensation, offline compensation, and
Biexponential data display for better visualization of digital data. All
experiment data, including instrument settings, plots, compensation matrices,
and statistics, is stored in a database for reuse and reanalysis.
- Multilaser multiparameter analysis cytometer
- Three lasers, 11 colors
- Automatic compensation (compensation controls are still needed)
- 355 nm (UV): BUV737, BUV395, DAPI
- 405 nm (Violet): CFP, Alexa Fluor 430, Pacific Blue, Marina Blue, Alexa Fluor 405
- 488 nm (Blue): FITC, PE, PerCP, Pe-Cy5 (aka CyChrome), PI, PerCp-Cy5.5, PE-Cy7, GFP, PE-Texas Red
- 633 nm (Red): APC, APC-Cy7, Alexa Fluor 647, Alexa 680
- Availability of UV gives a capability to perform Ca 2+ flux experiments.
- Unlike a FACSCalibur, it is possible to use PE-Cy5 and APC together.
BD Biosciences LSR II Flow Cytometer Specifications
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Performance
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Fluorescence Sensitivity
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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
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Fluorescence Resolution
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Coefficient of variation PI: Area of <3%, full G0/G1 peak for
propidium iodide (PI)-stained chicken erythrocyte nuclei (CEN).
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Fluorescence Linearity
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Doublet/singlet ratio of 1.95–2.05 for CEN stained with PI and
excited with the 488-nm blue laser
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Forward and Side Scatter Sensitivity
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Enables separation of fixed platelets from noise.
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Forward and Side Scatter Resolution
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Scatter performance is optimized for resolving lymphocytes,
monocytes, and granulocytes.
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Data Acquisition Rate
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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.
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Excitation Optics
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Excitation Optical Platform
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The BD LSR II optical layout allows for up to 4 laser wavelengths for
laser excitation.
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Laser Wavelengths and Laser Power
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355 nm: 20 mW 405 nm: 25 mW 488 nm: 20 mW 640 nm: 40 mW
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Optional Solid-State Lasers (available as a custom option)
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New options are developed on a regular basis. Please check with your
local sales representative for the latest custom options.
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Flow Cell Design
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Rectangular Quartz Cuvette: Internal cross-section, 430 x 180 mm
External quartz cuvette surfaces are antireflection coated for maximum
transmission of laser light. Fixed optical assembly of the lasers, with up to
4 spatially-separated laser beams.
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Emission Optics
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Optical Coupling
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The quartz cuvette is coupled to the emission lens by a
refractive-index– matching optical gel for optimum collection efficiency.
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Forward Scatter Detection
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Photodiode detector with a 488/10 bandpass filter
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Side Scatter Detector
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Photomultiplier tube (PMT) with a 488/10 bandpass filter
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Emission Optical Design
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The BD LSR II uses BD’s patented octagon and trigon detection system.
Using reflective optics and fiber-coupled PMTs ensures more efficient light
collection than that obtained from transmission optics. Please see the
separate filter guide for information on dye and filter options.
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Fluidics
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Sample Flow Rates
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Front key panel provides three modes: RUN, STNDBY, and PRIME
Continuously adjustable flow rate, plus three preset flow rates: LO = 12
µL/min MED = 35 µL/min HI = 60 µL/min
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Standard Fluidic Reservoirs
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One 8-L sheath and 10-L waste container provided.
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Recommended Fluidics Upgrade Option
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BD FACSFlow™ supply system: automated fluidics system, which includes
a rolling cart and two 20-L Cubitainer® packages.
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All of our equipment is fully tested to perform at factory
specifications before being shipped.
Included with the BD Biosciences LSR II Flow Cytometer: Includes: > HP Z220 Data Station > FacsDiva Software > Full 90 Day Parts and Labor Warranty BD LSR II Flow Cytometer Manual
 Click Here to View the Technical Specs for the LSR II
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