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BD Accuri C6 Cytometer Flow cytometry within reach

Flow cytometry within reachThe BD Accuri C6 is a personal flow cytometer that brings cell analysiswithin reach by being easy to use, simple to maintain, and affordable.The analytical power and versatility of today’slaser-based flow cytometry systems haveunlocked the mysteries of cell biologyand empowered entirely new fields ofresearch. As a result, flow cytometry hasbecome a staple of modern laboratoriesaround the world. Innovations in ease of usereflected in the BD Accuri C6 cytometer makethese powerful capabilities more accessibleto a new generation of flow cytometry users.The compact footprint and transportableweight of the BD Accuri C6 also make ita valuable personal use tool for experiencedresearchers who want a cytometer to beeasily available when and where theyneed it.Many BD Accuri C6 cytometer users canbegin collecting and analyzing data withthe help of a quick start guide. The intuitiveinterface of the software guides the userthrough workflows. A wide dynamic rangeof over 7 full decades ensures that all datais available all of the time. Informationobtained from the BD Accuri C6 can bere-analyzed at any time if gating orcompensation changes are required, or toaccommodate new research.The BD Accuri C6 is small enough to easilyfit on a benchtop and can be placed in alaminar flow hood if biohazard containmentis required. It measures 11 x 14.75 x 16.5inches (H x W x D) (27.9 x 37.5 x 41.9 cm)and weighs just 30 pounds (13.6 kg).3

Pre-optimized detectors,minimized setup timeThe system is equipped with a blue and red laser, two lightscatter detectors, and four fluorescence detectors withoptical filters optimized for the detection of FITC, PE, PerCP,and APC. A compact optical design, fixed alignment, and preoptimized detector settings make the system easier to use.Optional filters and the Selectable Laser Module expand theavailable fluorochrome combinations. During manufacture,laser and optical alignments are set and locked down. Theresult is that each BD Accuri C6 cytometer is manufacturedwith standardized fluorescence performance so that usersdo not need to adjust detector voltages.Data is digitally collected over a wide dynamic range of7.2 decades (16 million channels of digital data), makingall data available to users as needed. Gating strategies andfluorescence compensation values can be set before, during,or after data collection. After data is collected, theBD Accuri C6 software Zoom function allows visualizationof data at any scale, so that users can precisely set gatesand regions.Should updates in the values be required later, or if optimization is needed, simply change the settings and re-analyzethe data. This flexibility also allows data to be re-examinedto accommodate new research findings.The system has been put through intense testing to ensurethat the design can withstand rugged conditions. Providedthe system is anchored, it can run samples even if thebenchtop is in motion, for example, onboard a ship.A06 CD4 CD8 CD3Gate: (P1 in all)10 610 6A06 CD4 CD8 CD3Gate: (P1 in all)PE CD4-A10 310 410 510 110 1.710 310 410 5CD3 CD4 33.6%10 210 410 310 1.7PE-Cy7 CD8-A10 5CD3 CD8 7.1%10 1.710 610 310 6300V1-R84.5%A07 CD45 CD4 CD8 CD3Gate: (CD3 CD4 in (P1 in all))V2-LV2-RCount100Count50200V1-L15.5%10 210 310 4FITC CD45RA-A410 50After FITC staining, subpopulations of CD45RA and CD45RA–cytotoxic and helper T cells were identified.A07 CD45 CD4 CD8 CD3Gate: (CD3 CD8 in (P1 in all))0Thawed human peripheral blood mononuclear cells (PBMCs)were stained with directly labeled anti-CD45RA FITC, CD4 PE, CD8PE-Cy 7, and CD3 APC in PBS 1 mg/mL of BSA, covered, on ice,for 30 minutes. Cells were acquired and gated on lymphocytes toidentify CD3 CD8 cytotoxic (blue) and CD3 CD4 helper (red) T-cellpopulations.100T-cell phenotyping, 4-color analysis10 4APC CD3-AAPC CD3-A10 510 610 210 310 4FITC CD45RA-A10 510 6

A10 6.810 6.8 10 6.810 610 610 5.110 610 610 710 710 7JC-1 FL-2-AJC-1 FL-2-A10 6P10P10P1010.6%10.6%10.6%10 510 610 510,000,00014,540,96910 4.6 10 4.610 50 540,96914,540,969 10 4.6P9P9 P912.9%12.9%12.9%10 5.110 5.110 610 5.1JC-1 FL-2-AP9P9 CCCPJC-1 JC-1Gate:(P1ininall)Gate:Gate:(P1(P1all)in all)JC-1 FL-2-A10 710 710 tedJC-1 JC-1Gate:(P1ininall)Gate:Gate:(P1(P1all)in all)10 600P1P1 P190.0%90.0%90.0%CJC-1 FL-2-AJC-1 FL-2-A10 C-A2,000,0002,000,000The BD Accuri C6 can perform most flow cytometricapoptosis assays, including Annexin V, caspase activation,PARP cleavage, mitochondrial membrane potential (ΔΨm),and DNA fragmentation. (A) K562 cells (human chronicmyelogenous leukemia) were treated with CCCP to induceapoptosis, then stained with JC-1 according to theBD MitoScreen (JC-1) protocol (Cat. No. 551302). Thecells were washed and collected on the BD Accuri C6.CCCP treatment (C) resulted in a shift in ΔΨm (red togreen) vs untreated cells (B), indicating apoptosis.00Apoptosis teduntreatedJC-1 JC-1Gate:[NoGating]Gate:Gate:[NoGating][No Gating]10 5.110 5.1A10 4.610 510 4.6 10 4.610 5P10P10P1084.0%84.0%84.0%10 510 610 610 6JC-1FL-1-AJC-1JC-1FL-1-AFL-1-A10 6.810 6.8 10 6.85

Sample flexibility with optionalwalkaway sample loadingA unique low-pressure pumping system drives the fluidics.A sheath-focused core enables event rates of up to 10,000events per second and a sample concentration of over5 x 106 cells per mL. In addition, the system derivessample volume and can calculate absolute counts or sampleconcentration per microliter.The non-pressurized system supports any brand of 12 x75-mm (or smaller) sample tubes, including microcentrifugetubes and tubes made of polypropylene or polystyrene. TheBD Accuri C6 cytometer simplifies system maintenance withautomatic cleaning cycles on instrument shutdown. Thesystem can employ laboratory-grade water for sheath fluid,reducing operating costs.For walkaway convenience, the optional BD CSampler accessory offers reliable and easy-to-use automation.The system supports 48- and 96-well plates and deepwell plates, and is also supplied with a 24-tube rack forstandard 12 x 75-mm tubes. They are processed directly inthe BD Accuri C6 cytometer, saving time. The BD CSampleradds minimal footprint to the BD Accuri C6, about threefeet square for the pair, keeping the benchtop free forother uses.To streamline sample processing, the BD CSampler allowsmultiple collection settings to be applied to plate or tuberuns. To process a sample immediately, a run can be pausedusing the Interrupt function. When the priority operationis complete, the original plate can be returned to theBD CSampler to resume the original run. Easy-to-readsoftware messages keep users informed of system status.A: Beckman Coulter Cyan ADPA. Beckman Coulter Cyan ADPGap-free analysis of intracellular calcium1031031031021021011021011000 secData from Vines A, McBean GJ, Blanco-Fernández A. A flow cytometric method for continuous measurement of intracellular Ca2 concentration. Cytometry Part A. 2010;77:1091-1097;reproduced courtesy of the authors.488: 530/40: Fluo-4104488: 530/40: Fluo-4B The three right-hand cytograms show data obtained on aBD Accuri C6, adding the same compounds in open Eppendorftubes without interrupting sample acquisition. No time gaps wereobserved; otherwise, both methods obtained comparable data.104488: 530/40: Fluo-4A The three left-hand cytograms show calcium data obtained ona Beckman Coulter CyAn ADP using the “stop-flow” method,showing time gaps when control and test compounds were added.1042 min 30 sec 5 min 7 min 30 sec 10 minTime1001010 secA231872 min 30 sec 5 min 7 min 30 sec 10 minTime1000sA23187T7.2106105o-407.2106Gate: [No Gating]105o-40Gate: [No Gating]1005o-466107.2B. BD Accuri C6

Applications in kinetic analysis of cellular responsesChanges to intracellular calcium (Ca2 ) levels can occurrapidly, in some cases within nanoseconds of stimulation,and obtaining accurate data is a significant researchchallenge. To add test compounds to the cell suspension,a “stop-flow” method is often used in which samplingis paused, the sample tube opened, the agonist added,and the tube resealed. This technique leaves a gap in datacollection that may miss essential changes in Ca2 levels.The BD Accuri C6 employs non-pressurized peristaltic pumpsin an open fluidics system. Open tubes allow convenientaddition of test compounds to the cell suspension withoutinterrupting sampling. This “continuous-flow” methodenables non-stop analysis of calcium flux and other kineticcellular responses, such as pH, reactive oxygen andnitrogen species, mitochondrial membrane potential, andnanoparticle ulterCyanCyanCyanADPADPADP101 101 101102 102 102101 101 101100 100 100488: 530/40: Fluo-4102 102 102488: 530/40: Fluo-4102 102 102488: 530/40: Fluo-4103 103 103488: 530/40: Fluo-4103 103 103488: 530/40: Fluo-4103 103 103488: 530/40: Fluo-4104 104 104488: 530/40: Fluo-4104 104 104488: 530/40: Fluo-4104 104488: 530/40: Fluo-4104101 101 101100 100 100100 100 1000 sec0 sec20minsec2 min30 2sec30minsec530minsec5 min75minmin7 min30 7sec30minsec1030minsec10 min10 min0 sec0 sec20minsec2 min30 2sec30minsec530minsec5 min75minmin7 min30 7sec30minsec1030minsec10 min10 min0 sec0 sec20minsec2 min30 2sec30minsec530minsec5 min75minmin7 min30 7sec30minsec1030minsec10 min10 hapsigarginA23187A23187A23187B: BD Accuri C65m 5m0.0s0.0s5m 0.0sTimeTimeTime5m 5m0.0s0.0s5m 01011021031041010101011010345488:2 530/40:Fluo-4107.2488: 530/40: Fluo-41488:102 0.0s 0.0s0.0s0.0s 0.0sA23187A23187A23187 Gating]106107.2106107.21061010101011010345488:2 530/40:Fluo-4107.2488: 530/40: Fluo-41488:102 00.0s0.0s 0m10m0.0s10m0.0s 0.0s0.0s0.0s 0.0s5m 5m0.0s0.0s5m 0.0sTimeTimeTime10m10m0.0s10m0.0s 87A23187Gate: [No Gating]106107.215 min 7 min 30 sec 10 minTime102 min 30 secThapsigargin A23187100 sec345488:2 530/40:Fluo-4100110110488: 530/40: Fluo-4102101488:102 530/40:103104Fluo-4105488: 530/40: riC6C6C610405o-4ec 10 minFLUIDICS7