Let the BD FACSAria II
accompany your best performance everMeet the new BD FACSAria II
high-speed, fixed-alignment cuvette cell sorter.
The first generation BD FACSAria™ system brought the complex worldof cell sorting to a broader audience of researchers. Now the new
BD FACSAria II system makes cell sorting even easier and moreaccessible.
Designed with input from customers around the world, the new
BD FACSAria II improves ease of use, flexibility, and aseptic capability
to make it simply the best choice for consistent results in sorting acrossa broad range of applications.
The BD FACSAria II incorporates the unique fixed alignment
technology that revolutionized cell sorting over five years ago
with the introduction of the BD FACSAria. Fixed alignment
minimizes startup time and improves reproducibility. Gel coupling
to the objective lens improves collection efficiency and increases
sensitivity and resolution needed for multicolor sorting applications.
The BD FACSAria II advances cell sorting with multiple improvements including:
New fluidics and nozzle designs
A new fluidics design improves aseptic setup
and cleaning. New nozzle sizes enable awide range of particles to be sorted andnew nozzle design allows for reproducible
insertions. Software wizards and automation
simplify aseptic sort setup, drop delay
determination, and clog detection.
Better workflow and productivity
The BD FACSAria II also raises the bar onease of use to improve both workflow andproductivity with improvements across theplatform. For example, a universal sort tube
collection mount now makes it easier toinsert and remove tube holders. In addition,
software improvements dramatically cut thetime spent on cytometer setup, adjustment,
and maintenance.

New 375-nm Near UV laser
The 375-nm Near UV laser option isavailable to expand the range of application
choices. The beam geometry is optimized formost hematopoietic stem cells. The top hat
design of the 375-nm Near UV laser triples
the amount of light at the sample intercept.
The 375-nm Near UV laser can be operated
together with the red and blue lasers.
Upgrades from BD FACSAria to
BD FACSAria II
Packages will be available to field upgrade
BD FACSAria instruments to the capabilitiesof the BD FACSAria II.

Discover what makes the BD FACSAria II theplatform of choice worldwide—then let it
accompany your best performance ever.

FLUIDICS

First II Deliver

New fluidics design enables easy
aseptic setup and effective cleaning.
The fluidics system moves particles from thesample injection chamber through the cuvetteflow cell for interrogation and sorting into acollection device.
Aseptic setup and cleaning are achieved througha new fluidics design that is both effective
and easy to use. It features integrated valve
manifolds, and a streamlined fluidics path,to allow for improved aseptic sort setup. In
addition, new software wizards make aseptic
sort setup easy.

Fluidics cart

flow cellinterrogation point
laser beams
nozzlenozzle locking
lever

Cuvette flow cell
Hydrodynamic focusing within the flow
cell forces particles through the cuvettein a single-file stream where laser lightintercepts the stream at the sample
interrogation point. The unique flow cell
design permits particles to flow throughthe cuvette at a low velocity, allowing
longer exposure to laser energy. This
ensures optimal resolution and sensitivity
even at high speed sort settings. After
passing through the cuvette, the stream
accelerates through the nozzle and droplets
are formed for sorting.

The heart of a flow cytometer—
the gel-coupled cuvette flow cell
The gel-coupled cuvette flow cell is where the sample
is interrogated and measurements are performed. The
BD FACSAria was the first cell sorter with a flow cellin true fixed alignment with the laser and gel coupled
to the collection optics. Fixed alignment of the flow
cell and laser helps to ensure that the laser is precisely
focused on the sample stream and that each of thelasers can generate the greatest amount of signal.
Fixed alignment minimizes startup time, improves
experiment-to-experiment reproducibility, and enables
automated daily quality control. Gel coupling tothe objective lens improves collection efficiency andincreases sensitivity and resolution needed for multicolor applications. This design enables multicolor
sensitivity even at high-speed sort settings. Since
particle interrogation occurs above the nozzle, insertionand removal of the nozzle can occur without realigning
the optics or the fluid stream.
The Fluidics Cart
A separate fluidics cart supplies sheath and cleaning
fluids and collects waste from the cytometer. The selfcontained fluidics cart supplies the air pressure andvacuum needed to achieve pressure from 5 to 75 psi
to accommodate a variety of cell sorting applications.
Air pressure is adjusted within BD FACSDiva™ software.
The fluidics cart holds a 10-L stainless steel sheathtank, a 5-L stainless steel ethanol shutdown tank, a
10-L waste container, and three 5-L auxiliary cleaning
fluid containers. The stainless steel sheath tankcan be autoclaved for aseptic cleaning. The cart istypically positioned directly under or to the left of thecytometer.

Cuvette flow cell and nozzle

BD Accudrop Technology
Patented BD Accudrop™ technology makes
it easy to determine an accurate dropdelay value. Drop delay determination is
now easier due to software automation.

Software display of stream breakoff
After the drop delay is calculated, thesystem automatically adjusts to maintain aconstant breakoff, dubbed the Sweet Spot.
If a clog is detected, the system stops thesort and protects the collection tubes.

SAMPLE TO SORT C

Committed II Advance

Aerosol management isengineered into the system.
After leaving the nozzle, particles pass throughthe sort block where they are deflected into acollection device in the sort collection chamber.
The sort block houses the high-voltage
deflection plates used to deflect side streams
during sorting. A novel deflection plate design
fixes plates in position for more efficient andreproducible deflection.
The aspirator and aspirator drawer are also
housed in the sort block. The aspirator drawer
keeps the sort collection tubes covered until
sorting begins and automatically closes toprotect sort collection tubes when the Sweet
Spot is on and a clog is detected.

“The BD FACSAria is the fastest, most sensitive cell
sorter that
I have worked with, and the simple design ofthe new fluidics system makes sterile sort setup abreeze.”COLLECTION

Aerosol management
Engineered with aerosol management in mind, theBD FACSAria II features an enclosed pathway from thesample injection chamber to the sort collection tubes.
For an added level of aerosol management, theBDTM Aerosol Management Option (AMO) evacuates
the sort collection chamber and traps aerosolized
particles during sorting.
Sample injection chamber
The sample injection chamber is where the sample isintroduced into the flow cytometer. During acquisition,
the chamber is pressurized, forcing the sample to the
cuvette flow cell. Sample injection chamber temperature
control and agitation are standard with the
BD FACSAria II cell sorter. The BD FACSDiva software
controls both sample agitation and temperature
settings.
After a sample tube is run, both the inside and outsideof the sample injection tubing are flushed to minimize
carryover. A variety of tube holders are provided, from
15-mL centrifuge tubes to 1.0-mL microtubes. New 35and 50-micron sample line filters minimize clogging
during a sort.
Sort collection chamber
The new universal mount in the sort collection
chamber makes inserting the tube holders easier.
This design is more robust and helps maintain aseptic
conditions. Temperature control for both sort collection
tubes and plates is available as an option.

Sort collection chamber

Nozzle selection and design
An expanded set of nozzles lets users sorta wide range of particle sizes. Four nozzle
sizes are supported: 70, 85, 100, and 130
microns. An improved nozzle insertion
design offers tight registration for asecure fit. This means a reproducible drop
profile after every insertion, allowing forreproducible instrument setup. Nozzles
are readily accessible and easy to change.
Software enhancements include a nozzlespecific sort setup feature that matchespressure settings to the nozzle in use.

OPTICS
Innovative optics design increases
multicolor detection and efficiency.

More II See

Innovative designs for both the excitation optics
and collection optics reduce excitation losses
and dramatically improve collection efficiency.

Excitation optics
The excitation optics consist of lasers, fiber optic cables,
beam-shaping prisms, and an achromatic focusing lens.
Fiber optics precisely direct the laser light into beamshaping prisms that transmit the laser light into a
focusing lens. The lens focuses the laser light into thegel-coupled cuvette flow cell. Since the optical pathway
and the sample core stream are fixed, alignment isconstant from day to day.

The optical design allows for the use of lower
powered, air cooled, and solid state lasers
for efficiency without sacrificing sensitivity
or resolution. No special power or cooling isneeded.

Collection optics
The collection optics are set up in octagon- and trigonshaped arrays that maximize signal detection from eachlaser. This is accomplished by transmitting the highest
wavelengths to the first photomultiplier tube (PMT), andreflecting lower wavelengths to the next PMT througha series of longpass dichroic mirrors. Bandpass filtersin front of each PMT allow fine-tuning of the spectral
wavelengths that need to be collected.
This design is based on the principle that light reflection
is more efficient than light transmission. Emitted lighttravels to each PMT detector via reflection and istransmitted through only two pieces of glass to reachthe detector. This greatly increases sensitivity andresolution for each color in a multicolor panel.

Aria II CD4 APC

200

200

200

150

100

SSC-A (x 1,000)

250

50

150

150

100

100

50

0

102

-308

103

104

105

50

0

103

104

105

0

Canto II CD4 APC

200

200

200

50

150

102

103

FITC-A

104

105

100

50

0
-110

105

150

100

50

0
-40

104

SSC-A (x 1,000)

250

100

103

Pacific Blue-A

Canto II CD4 Pacific Blue

250

150

102

-228

CD4 APC-A

250

SSC-A (x 1,000)

SSC-A (x 1,000)

102-1,328

CD4 FITC-A

Canto II CD4 FITC

FACSCanto II

Aria II CD4 Pacific Blue

250

SSC-A (x 1,000)

FACSAria II

SSC-A (x 1,000)

Aria II CD4 FITC
250

102

103

APC-A

104

105

0
-39

102

103

104

Pacific Blue-A

105

Platform comparison using CD4
Comparison of whole blood stained withsingle-color CD4 FITC, CD4 APC, and CD4
Pacific Blue™ and run on both the
BD FACSAria II and BD FACSCanto™ II
systems. The BD FACSAria II was set for
high-speed sorting (70 psi and 90 kHz)and both instruments were set up using
BD Cytometer Setup and Tracking software.

Excitation optics focus the 488-nm, 633-nm, and 405-nm laser beams

streamcuvette flow cell
individual collection
fibersnozzlefluorescence objective lensGel-coupled cuvette flow cell andfluorescence objective lens assembly
The cuvette is gel coupled to thefluorescence objective lens to transmit thegreatest amount of emitted light from theinterrogation point to the collection optics.
The lens focuses fluorescent light emitted
at each of the laser interrogation points
onto individual collection fibers. Thesefibers transfer the emitted light to thecollection optics.

OPTICS

Able II Perform

Detects up to 15 parameterssimultaneously for multicolor flexibility.

For many users, the capability of an advanced
cell sorter is defined by its flexibility, which inturn is defined by the number of parametersthat can be detected simultaneously. To provide
maximum flexibility, the BD FACSAria II cell
sorter can be configured with up to four aircooled, solid state lasers and can detect up to
15 parameters simultaneously—13 fluorescentmarkers and two scatter parameters when
fully equipped.

A standard system is equipped with an octagon
containing six PMTs that detect light from the488-nm (blue) laser, and a trigon containing two
PMTs that detect light from the 633-nm (red) laser.
The third laser option adds an additional trigon
containing two PMTs to detect light from the405-nm (violet) or 375-nm (Near UV) lasers. Standard
instruments can be configured with all four lasers.
375-nm Near UV Laser
The 375-nm Near UV laser option is available to expand
the range of application choices. Measuring only
19 × 7 × 7 cm, the fully-housed 375-nm laser fits in thesmall space immediately left of the flow cell. The laser
produces a 10-mW focused beam to illuminate the cells
in the flow cell without the aid of fiber optics.

375-nm Near UV laser
The top hat design of the 375-nm Near UV
laser modifies the usual Gaussian beam(in red) that illuminates a sample at higher
intensity in the middle of the beam(in blue) and lower intensity at the edges.
The composite “square” beam illuminates
the middle and edges of the sample corestream at the same intensity. The “square”
beam almost triples the amount of light at
the sample intercept.

1
0.9

sample core stream

0.8

Intensity

0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
-50

-40

-30

-20

-10

0

10

Guassian (red line) vs. Top Hat (blue line)
illumination

20

30

40

50

Trigon and octagon detector arraysD(x 1,000)250

450/20-A

B

150

Mouse stem cell side population detection
As a consequence of the even sample
illumination, cells have a greatly improved
coefficient of variation (CV).

200

F

H
100

G

50

A
50

100
150
675 LP-A

200
250(x 1,000)

ECTransmission pathways in an octagon

CONTROLS AND

Easy II Handle

Compatibility with other BD analyzers
and sorters makes it easier to move fromanalysis to cell sorting.

The BD FACSAria II cell sorter places instrument
setup, QC, sorting, and data management
under the control of BD FACSDiva software
so researchers can spend more time on
experiments.

Digital electronics
The BD FACSAria II cell sorter uses digital electronics toacquire data at rates up to 70,000 events per second
for up to 8 parameters, with linear results over a broad
dynamic range.
Cytometer Setup and Tracking software
Enhancements to BD FACSDiva software include a
new Cytometer Setup and Tracking (CS&T) feature that
automates flow cytometer setup, adjusts for instrument
variability, and establishes baseline settings. Chancesof operator error are reduced, and results are moreconsistent.
The setup features let users define a configuration
baseline and run a performance check. The software
will then automatically set up the cytometer to thisestablished baseline. To match the exact baseline, the
CS&T software automatically adjusts for reproducible
performance of the instrument. This ensures the greatest
possible consistency from one day to the next, saves
time, and improves reliability of experiment results.
The software also allows for the creation of applicationspecific settings to allow for rapid performance ofroutine experiments in a more consistent manner.
Tracking capabilities in the software measure a numberof instrument settings and report on performance.
Levey-Jennings plots help users understand instrument
performance and identify maintenance issues.

Fast startup to sort timeu Turn on the sorter.
v Start the fluidics.
w Perform automated setup, QC,
and drop delay optimization.
x Optimize the sample.
y Perform the sort.

ANALYSISBD Cytometer Setup and Tracking software

Proccessed Events

Outperforms analog from 0 to 100,000 events/sec

90000
80000
70000
60000
50000
40000
30000
20000
10000
0

BD FACSAria II: uncompensated
BD FACSAria II: compensatedDead time of 5.5 µsec
BD FACSDiVa: compensated

0

30000

60000

90000 120000 150000 180000 210000

Event Rate

BD FACSAria II system acquisition rates

SERVICES

Ready II Deliver

Training
Hands-on training is included with each BD FACSAria II
cell sorter. In addition, training courses are held at
BD training centers worldwide. BD flow cytometry
training courses combine theory and practice toprovide participants with the skills and experience
they need to take full advantage of the capabilitiesof the BD FACSAria II cell sorter.

BD Biosciences is fully committed to thesuccess and satisfaction of its customers.
The BD FACSAria II cell sorter is backed by aworld-class service and support organization
with unmatched flow cytometry experience.

Technical application support
BD Biosciences technical applications support specialists
are available to provide field- or phone-based
assistance and advice. Expert in a diverse array of
topics, BD technical application specialists are wellequipped to address customer needs in both instrument
and applications support.

Since 1974, BD has innovated flow
cytometry for optimal performance,ease of use, and improved workflow. This
expertise is made available to BD FACSAria II
customers through comprehensive training,
applications and technical support, andexpert field service.

Field service engineers
When instrument installation or service is required,a BD Biosciences Technical Field Service Engineer canbe dispatched to the customer site. BD Biosciences field
service engineers are located across the world. On-site
service and maintenance agreements are available toprovide long-term support for the BD FACSAria II.
Custom Services
Mobilizing technology for research applications requires
close collaboration. The Custom Technology Team (CTT)at BD Biosciences works with customers to provide
solutions through custom reagents, panels, or assay
protocols.
Staffed by leading scientists with the breadth and depthof scientific and technical expertise, the CTT team will
coordinate with researchers to study the problem athand, make recommendations, and help implement thesolutions. In this way, BD Biosciences technical know-how
is translated into practical solutions that allow customers
to focus on research.
Special Order Research Products
Instruments can be customized to meet customer
requirements via the Special Order Research Products(SORP) programs.

Regional Offices
Asia Pacific

BD Singapore
Tel 65.6861.0633
Fax 65.6860.1593

bdbiosciences.com/offices
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Fax 800.325.9637
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bdbiosciences.com

BD flow cytometers are Class I (1) laser products. © 2008 Becton, Dickinson and Company. All rights reserved. No part of this publication may be reproduced, transmitted, transcribed, stored in retrieval systems, or translated into any language or computer language, in any form orby any means: electronic, mechanical, magnetic, optical, chemical, manual, or otherwise, without prior written permission from BD Biosciences.
The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc., for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for
Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
For research use only. Not for use in diagnostic or therapeutic procedures. Purchase does not include or carry any right to resell or transfer this product either as a stand-alone product or as a component of another product. Any use of this product other than the permitted use
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BD, BD Logo and all other trademarks are property of Becton, Dickinson and Company. © 2008 BD

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