BD FACSAria Flow Cytometer - HKU Paedpaed.hku.hk/services/lab_area/equipment/flow/Brochures.pdf ·...

BD FACSAria Flow CytometerThe first high-speed sorter with a fixed-alignment

cuvette flow cell from BD Biosciences

BD BiosciencesClontechDiscovery LabwareImmunocytometry SystemsPharmingen

SJ-0003-00Brochure 2/7/03 5:46 PM

1 www.bdbiosciences.com

From Pioneers to Revolutionaries inFlow Cytometry Instrumentation

BD Biosciences continues to providethe most advanced instrumentation inflow cytometry. Since 1974 when wedeveloped the first commerciallyavailable fluorescence activated cellsorter (FACS™) in collaboration withStanford University, BD Biosciences has been the leader in flow cytometrytechnology and innovation. BD Biosciencescontinues to be the industry leaderwith a revolutionary new high-speedcell sorter, the BD FACSAria™instrument. This novel instrumentplatform has been created from theground up with over 25 years of experience in flow cytometryinstrumentation. Our technologicaladvances in flow cytometry helpcustomers advance their research anddiscovery.

The BD FACSAria™ cell sorter is namedfor the musical term Aria, meaning a striking solo performance. The BD FACSAria cell sorter embodies thisname because it is able to performindependently without the burden of special room requirements or yearsof technical expertise. It is easy to operate and provides superiorperformance in high-speed cell sortingand multicolor analysis.

BD Biosciences is committed to helpingeach laboratory excel scientifically by providing the most advancedtechnology in flow cytometry. Thereliability and performance of ourinstrumentation, reagents, service, andsupport have been key to customersatisfaction. BD Biosciences offers thecomplete solution in flow cytometry.

SJ-0003-00Brochure 2/7/03 5:46 PM

Next Generation Flow Cytometry High-Speed Sorter

The BD FACSAria cell sorter sets a new standard for high-performance flow cytometry. Based on an entirely newdesign in instrumentation, the BD FACSAria instrument is the first benchtop sorter that incorporates a fixed-alignment cuvette flow cell. The numerous technological advances embodied in the BD FACSAria cell sorter reducethe cost of owning a high-speed sorter and accelerate research by providing unparalleled, easy-to-use, high-performance sorting and analysis.

2www.bdbiosciences.com

SJ-0003-00Brochure 2/7/03 5:47 PM

The Heart of the BD FACSAria Cell Sorter

The cuvette flow cell is the heart of the BD FACSAria cellsorter. New technologies incorporated in the instrumentdesign create a truly fixed-alignment optical path. Thefixed optical alignment frees the operator from tediousinstrument optimization, allowing the focus to center onthe science rather than the instrument.

The cuvette flow cell achieves high sensitivity in signaldetection with low-powered lasers, allowing air-cooledand solid state lasers to be utilized in this system. This, in turn, eliminates the need for special power and coolingrequirements associated with high-powered lasers used inconventional stream-in-air sorters.

Fiber optics direct and focus the laser light in a preciseand constant manner on the sample core stream. Fiberoptics steer the three lasers, 488 nm, 633 nm, and 407 nm,onto the alignment prisms, and then the lasers are focusedon the cuvette flow cell (see Figure 1). Because the place-ment of the sample core stream within the cuvette flowcell and the laser alignment are fixed, daily instrumentoptimization of the cytometer is no longer necessary.

Two major advantages of the cuvette flow cell design areimproved light excitation and collection optics. Excitationby the BD FACSAria cell sorter differs from a stream-in-airsorter. The sample passes through the laser beams at alower velocity, allowing longer exposure to laser energy.The stream is accelerated only as it enters the nozzle tip. The drop drive then breaks the stream into dropletsto be sorted.

Another advantage of the BD FACSAria cell sorter is howit collects light signals. Stream-in-air sorters can overcomeexcitation losses by increasing laser power. However, atsome point, most fluorophores saturate. Only a bettercollection system can increase sensitivity. A gel-couplingcuvette flow cell collects at least four times the amountof light over stream-in-air sorters. The BD FACSAriainstrument is the only high-speed sorter with a gel-coupledcuvette, thus improving sensitivity in signal detection (<125 Molecules of Equivalent Soluble Fluorescein [MESF]).*See Figure 10 on page 8 for a demonstration of the highsensitivity achieved with the cuvette flow cell.

*Using SPHERO™ Rainbow Calibration Particles RCP-30-5A


Figure 1Excitation optics focus the 488-nm,

633-nm, and 407-nm laser beams

SJ-0003-00Brochure 2/10/03 6:24 PM

Revolutionary Optics System

The cuvette flow cell is gel-coupled to the fluorescent objective lens to transmit the greatest amount of light to the collectionoptics (see Figure 2). The fluorescent objective lens collects andfocuses the fluorescent light emitted at each of the three laserfocal points onto individual collection fibers. The collection fibersthen transfer the emitted light onto the collection optics. Thecollection optics are engineered to achieve the greatest amount of signal detection from each laser. This is accomplished bytransmitting the highest wavelengths to the first PMT andreflecting the lower wavelengths to the next PMT through a series of longpass dichroic mirrors. Bandpass filters in front of each PMT allow fine-tuning of the collected signal. Sincereflection is more efficient than transmittance, this design greatlyincreases the multicolor detection capabilities of the instrument(see Figure 3).

The collection optics consist of octagon- and trigon-shapedcollection devices. The octagon-shaped collection device for the488-nm laser detects up to seven fluorescent colors plus side scatter.The trigon-shaped collection devices for the 633-nm and 407-nmlasers each collect up to three fluorescent colors. With the additionof FSC, a total of 15 independent signals can be acquired at onetime. This entirely new optical system increases the quality andquantity of the information acquired from each sample.Representative data is shown in Figures 11 and 12 on page 8.


Cuvette

Stream

Lens

Individual collection

fibers

Figure 3Octagon-shaped collection optics

Fiber

A

H

F

DB

C

E

G

Figure 2Gel-coupled cuvette flow cell and

fluorescent objective lens assembly

SJ-0003-00Brochure 2/7/03 5:47 PM

Technological Advances in the Fluidics System

The fluidics system is another key technological advancementembodied in the BD FACSAria™ system. The fluidics systemincludes an entirely new means of delivering both the sample and the sheath fluid to the instrument. A new fluidics cartsupplies the sheath and cleaning fluids from non-pressurizedcontainers and collects the waste from the instrument.

The self-contained fluidics cart supplies air pressure and vacuumeliminating the need for an external source. This feature allowsthe instrument to operate in any lab or room without the specialand costly facility requirements of a traditional high-speed flowcytometer. The air pump provides pressure from 2–75 psi (14–517 kPa)to accommodate a variety of sorting applications. The fluidics cartholds four 10-L containers, two sheath and two waste, and three5-L auxiliary cleaning fluid containers (see Figure 4). The cart alsoaccommodates BD FACSFlow™ sheath fluid containers.

The fluidics system transfers sheath fluid from the fluidics cart into a plenum reservoir inside the BD FACSAria system (see Figure 5). The plenum reservoir holds the sheath fluid beforeit is pumped into the cuvette flow cell and removes any pressurechanges as the plenum reservoir is refilled. Thus, pressure is nolonger dependent on the level of fluid in the sheath container,and the reservoir removes small air bubbles from the sheathsupply. The entire fluidics system is designed to further enhance this easy-to-use instrument.

The instrument has a specialized sample injection chamber that accommodates different tube sizes to provide flexibility in experimental design (see Figure 6). Once a tube is placed in thetube holder of the sample injection chamber, the entire chamberbecomes pressurized. This allows even cracked tubes to be sampledand aids sample containment. The tube holder agitates the sample,keeping cells constantly suspended throughout a long sort.

Because the sample injection chamber is self-contained,specialized cleaning modes are available. After each tube isremoved, sheath fluid flushes the sample tubing inside and out to eliminate potential sample carryover. Using a special cleaningmode, the entire sample injection chamber can be filled with ethanol.


Figure 4Fluidics cart

Figure 5Plenum reservoir

Figure 6Sample injection chamber

SJ-0003-00Brochure 2/7/03 5:47 PM

New Era for High-Speed Cell Sorting

With the BD FACSAria cell sorter, high-speed sorting hasnever been easier to set up and perform. Advances ininstrument design give the BD FACSAria system improvedsort performance without sacrificing results. Due to theunique design of the cuvette flow cell, drop drivedisturbances that create noise on stream-in-air systemsare not seen on the BD FACSAria cell sorter. Unlikestream-in-air systems, there is no perceptibleperformance degradation of any parameter while usingthe drop drive. Thus, data quality is not compromisedduring sorting.

The nozzle tip is keyed into a fixed position at the end ofthe cuvette ensuring reproducible stream alignment intothe waste aspirator (see Figure 7). Fixed nozzle alignmentensures the stream will return to the same spot after anozzle tip is removed and reattached.

The 70- and 100-micron nozzle tip sizes are designed to accommodate most cell types. The nozzle sizes enablesorting at a variety of pressures and speeds. Two- andfour-way sorting are standard features available for a variety of bulk collection devices to accommodatedifferent tube sizes (for example, microtubes, 12 X 75 mm,and 15 mL). The optional automated cell deposition unit(ACDU) sorts into multiwell plates and onto microscopeslides (see Figure 14 on page 10). The flexibility of the BD FACSAria cell sorter provides the total solution for flow cytometry sorting.

Figure 7Cuvette flow cell and nozzle tip

SJ-0003-00Brochure 2/7/03 5:47 PM

Advanced Digital Electronics Achieve Unparalleled Performance

Advanced digital electronics improve sort performanceand provide significant advantages in instrument ease-of-use. The electronics eliminate dead time. This improvesthe number of signals that can be processed and sorted,thus increasing sort yield by reducing the number ofelectronic aborts. The electronics simplify compensationof all fluorescent signals with no limitation of inter- andintra-beam compensation.

Faster signal processing enables acquisition rates of up to 70,000 events per second for 15 parameters. The BD FACSAria system can achieve faster acquisition ratesthan an analog system. See Figure 8 for eight-parameterdata generated for instrument comparison.

Aerosol Management

While sample is acquired and sorted, the sample injectionand sort collection chambers are enclosed to help withaerosol management. The sort collection chamber iscompletely sealed and operates under negative pressurefrom the waste aspirator. To reduce the chance ofbiohazard exposure, the instrument rinses the sampletubing between samples.

The BD™ Aerosol Management Option (AMO) (see Figure 9)is an optional device that provides additional aerosolmanagement and removal within the sort chamber area.The AMO is equipped with a 0.01-µm pore size ultra-lowpenetrating air (ULPA) filter to trap aerosolized particles.The AMS evacuates the volume of the ACDU chamberand sort area 16 times per minute in normal evacuationmode and 69 times per minute in rapid evacuationmode.


Outperforms analog from 0 to 100,000 events/sec

Proc

cess

ed E

vent

s

Event Rate

8000090000

70000600005000040000300002000010000

00 30000 18000060000 90000 120000 150000 210000

BD FACSAria: uncompensatedBD FACSAria: compensatedDead time of 5.5 µsecBD FACSDiVa: compensated

Figure 9Aerosol Management Option

Figure 8BD FACSAria system acquisition rates

SJ-0003-00Brochure 2/7/03 5:47 PM


The BD FACSAria cell sorter achieves highseparation between negative and positiveevents as shown in Figure 12. In normalwhole blood stained with CD4 single-colorfluorophores and acquired at 70 psi and90 kHz, signal is detected from each laser:PE-Cy7 from the 488-nm laser, APC fromthe 633-nm laser, and Cascade Blue ® fromthe 407-nm laser.

The high sensitivity of the BD FACSAria cell sorter is shown in Figure 10. In DNA stained with PI and Hoechst at 70 psi and 90 kHz,CEN stained with PI resulted in a CV of 1.5% and with Hoechst a CVof 3.5%.

CENs

Co

un

t10

020

00

300

400

500

600

700

800

900

PI-A

CENs

Co

un

t25

050

00

750

1,00

0

200 25050 100 150

Hoechst-A200 25050 100 150

P1 P2

P1 P2

200

250

5010

015

0

200 25050 100 150FSC-A

SSC-

A(x

1,0

00)

(x 1

,000

)

(x 1,000)

105102 103 104

PE-Cy7-A

PE-Cy7

SSC-

A

APC-A

SSC-

A(x

1,0

00)

(x 1

,000

)

105102 103 104

Cascade Blue-A

Cascade Blue

APC

SSC-

A20

025

050

100

150

200

250

5010

015

020

025

050

100

150

105102 103 104

Figure 10DNA stained with PI and Hoechst

ZcYellow105102 103 104

105

102

103

104

HcRed

Fluorescent Proteins

Am

Cya

n

105102 103 104

105

102

103

104

Fluorescent Proteins

DsR

ed

Its revolutionary optics allow the BD FACSAria instrumentexpanded possibilities in multicolor analysis. In Figure 11, fourunique fluorescent proteins from BD Clontech are excited by the 488-nm, 633-nm, and 407-nm lasers: AmCyan, DsRed, ZcYellow,and HcRed.

Figure 11Fluorescent proteins

Figure 12Whole blood stained with CD4

single-color fluorophores

Representative Data

SJ-0003-00Brochure 2/7/03 5:47 PM

Increased Sorting Advances Through Software

BD FACSAria™ software provides unparalleled instrumentcontrol for flexible sorting and data analysis. All instrumentand sorting controls are embedded in the software tosimplify instrument setup and operation. New featuresprogrammed into the software provide the completesolution for high-performance cell sorting.

Fixed-nozzle alignment and a software breakoffmonitoring system ensure that the drop delay value does not change unless a nozzle is replaced. IntegratedBD FACS™ AccuDrop technology allows the user todetermine an accurate drop delay value. See Figure 13 fora software display of stream breakoff and side streams.

Comprehensive sort monitoring and clog detection are standard integrated software features that make the BD FACSAria cell sorter easy to use. The breakoffmonitoring system—referred to as the sweet spot inrecognition of Dr. Richard Sweet who holds the patent on this technology—automatically adjusts the amplitudeto maintain the same breakoff value throughout a sort.The clog detection feature signals the instrument to stopthe sort and protect the collection tubes if the instrumentcannot maintain the stream breakoff.

Increased drop resolution provides accurate investigation of each sorted drop and results in better sort purity and yield. Each drop is investigated in 32 increments. Byexamining the sorted drops in smaller increments, eachsort decision is accurate and efficient. Increased resolutionincreases sort accuracy and performance by sorting onlythe drops of interest.


Figure 13Software display of streambreakoff and side streams

SJ-0003-00Brochure 2/10/03 6:24 PM

Header

11www.bdbiosciences.comUnless otherwise specified, all products are for Research Use Only.Not for use in diagnostic or therapeutic procedures. Not for resale.

BD FACSAria Workstation and Software

The BD FACSAria system operates on a PC-basedWindows® platform. The system includes a variety ofremovable storage devices and comes equipped with ahigh-resolution LCD flat-panel monitor. Ethernet is builtin for networking capabilities. A three-port FireWire® PCIadapter is also provided. A wireless keyboard and mouseprovide freedom of movement and flexibility whileperforming sample optimization and sort setup.

Unique Design

The BD FACSAria instrument was engineered to be spaceefficient and compact with reduced overall size comparedto conventional sorters. The instrument can be installedand operated in virtually any room without special facilityrequirements. The instrument can be set up on a typicallaboratory benchtop or table and requires only a standardelectrical outlet.

Large water-cooled lasers are no longer necessary to achievehigh fluorescent sensitivity and sorting performance. Thefluidics system is supported entirely by the self-containedfluidics cart. The BD FACSAria instrument is revolutionary in all aspects of its design and operation.

Figure 14ACDU option allows plate

and slide sorting

SJ-0003-00Brochure 2/7/03 5:48 PM

Argentina/Paraguay/UruguayTel 54.11.4551.7100 x106Fax 54.11.4551.7400

Australia/New ZealandAustraliaTel 61.2.8875.7000Fax [email protected] ZealandTel 64.9.574.2468Fax [email protected]

AustriaSCIENTIFIC SUPPORT

Tel 43.1.706.36.60.44Fax [email protected] SERVICE

Tel 43.1.706.36.60.01Fax 43.1.706.36.60.11

Belgium Tel 32.53.720.600Fax [email protected] SERVICE

Tel 32.53.720.550Fax [email protected]

BrazilTel 55.11.5185.9995Fax 55.11.5185.9895

Central America/CaribbeanTel 506.290.7318Fax 506.290.7331

ChileTel 56.2 460.0380 x21Fax 56.2 460.0306

ChinaTel 8610.6418.1608Fax 8610.6418.1610

ColombiaTel 57.1.572.4060 x244Fax 57.1.244.1363

DenmarkTel 45.43.43.45.66Fax [email protected]

East AfricaTel 254.2.341157Fax [email protected]

Eastern EuropeTel 49.6221.305.161Fax 49.6221.305.418

EgyptTel 202.268.0181Fax 202.266.7562

FinlandTel 358.9.88.70.7832Fax [email protected]

FranceTel 33.4.76.68.36.40Fax 33.4.76.68.35.06SCIENTIFIC SUPPORT

Tel 33.4.76.68.34.25Fax 33.4.76.68.55.71CUSTOMER SERVICE

Tel 33.4.76.68.37.32Fax 33.4.76.68.35.06

GermanySCIENTIFIC SUPPORT

Tel 49.6221.305.525Fax [email protected] SERVICE

Tel 49.6221.305.551Fax [email protected]

GreeceTel 30.210.940.77.41Fax 30.210.940.77.40

Hong KongTel 852.2575.8668Fax 852.2803.5320

HungarySzerenaTel 36.1.345.7090Fax 36.1.345.7093

IndiaTel 91.124.638.3566.77/3219Fax 91.124.638.3225

IndonesiaTel 62.21.577.1920Fax 62.21.577.1925

ItalyTel 39.02.48.240.1Fax 39.02.48.20.33.36

JapanFujisawa Pharmaceutical Co., Ltd.(Reagents from ImmunocytometrySystems & Pharmingen)Tel 81.6.6206.7890Fax 81.6.6206.7934

KoreaTel 822.3404.3700Fax 822.557.4048

MalaysiaTel 603.7725.5517Fax 603.7725.4772

MexicoTel 52.55.5999.8296Fax 52.55.5999.8288

Middle EastTel 971.4.337.95.25Fax 971.4.337.95.51

Netherlands Tel 31.20.582.94.24Fax [email protected] SERVICE

Tel 31.20.582.94.20Fax [email protected]

North AfricaTel 33.4.76.68.35.03Fax [email protected]

NorwayImmunocytometry Systems &PharmingenLaborel S/ATel 47.23.05.19.30Fax 47.22.63.07.51ClontechTel 47.22.95.59.59Fax 47.22.95.59.40

Peru/Bolivia/EcuadorTel 51.1.430.0323Fax 51.1.430.1077

PhilippinesTel 632.807.6073Fax 632.850.1998

PolandTel 48.22.651.53.00Fax 48.22.651.79.24

PortugalEnzifarmaTel 351.21.422.01.00Fax 351.21.422.01.10

Saudi ArabiaTel 966.1.26.00.805/806Fax 966.1.26.00.804

South AfricaTel 27.11.807.15 31Fax 27.11.807.19 53

SpainImmunocytometry Systems &PharmingenSCIENTIFIC SUPPORT

Tel 34.91.848.81.77Fax 34.91.848.81.05CUSTOMER SERVICE

Tel 34.902.27.17.27Fax 34.91.848.81.04ClontechTel 34.91.848.81.85Fax 34.91.848.81.04

SwedenTel 46.8.775.51.10Fax [email protected]

SwitzerlandSCIENTIFIC SUPPORT

Tel 41.61.485.22.91Fax [email protected] SERVICE

Tel 41.61.485.22.22Fax [email protected]

TaiwanTel 8862.2722.5660Fax 8862.2725.1768

ThailandTel 662.643.1374Fax 662.643.1381

TurkeyTel 90.212.222.87.77Fax 90.212.222.87.76

UKTel 44.1865.78.16.88Fax 44.1865.78.16.27SCIENTIFIC SUPPORT

Tel 44.1865.78.15.66

VenezuelaTel 58.212.443.6411 x248Fax 58.212.442.4477

West AfricaSobidisTel 225.20.33.40.32Fax 225.20.33.40.28

02/2003 SJ-0003-00

Asia PacificBD SingaporeTel 65.6861.0633Fax 65.6860.1590

United StatesBD BiosciencesClontechFax 650.354.0775Discovery LabwareFax 978.901.7493Immunocytometry SystemsFax 408.954.2347PharmingenFax 858.812.8888Customer/Technical ServiceToll free 877.232.8995www.bdbiosciences.com

JapanNippon Becton DickinsonToll free 0120.8555.90Tel 81.24.593.5405Fax 81.24.593.5761

Clontech Company(Clontech Products)Tel 81.3.5324.9609Fax 81.3.5324.9637

EuropeBelgiumTel 32.53.720.211Fax [email protected]

CanadaBD BiosciencesToll free 888.259.0187Tel 905.542.8028Fax [email protected]

The BD FACSAria™ flow cytometer is For Research Use Only. Not for use in diagnostic ortherapeutic procedures.

BD, the BD logo, FACS, FACSAria and FACSFlow are trademarks of Becton, Dickinson andCompany. Windows is a registered trademark of Microsoft Corportaion. Cascade Blue is aregistered trademark of Molecular Probes, Inc. SPHERO is a trademark of Spherotech, Inc.Firewire is a trademark of Apple Computer, Inc., registered in the US and other countries.©2003 BD

Visit us at BDBiosciences.com.

BD FACSAria Flow Cytometer - HKU Paedpaed.hku.hk/services/lab_area/equipment/flow/Brochures.pdf ·...

Documents

Transcript of BD FACSAria Flow Cytometer - HKU Paedpaed.hku.hk/services/lab_area/equipment/flow/Brochures.pdf ·...