Data prepared in collaboration with Juli Dyble and Gary Fahnensteil, NOAA Great Lakes Environmental Research Laboratory, Ann Arbor, MI, USA. The sophisticated digital signal processing system has a seven-decade dynamic range that encompasses almost the entire known span of plant C-values (haploid nuclear DNA content). Cell concentration can be calculated directly from BD Accuri™ software statistics tables without the addition of counting beads. Figure 7 Figure 8 Figure 9 7. Using the BD Stemflow™ Human MSC Analysis Kit (Cat. "Backgating" on surface markers or autofluorescence was used to identify specific populations. Cellular processes and events are probabilistic and take place in a context of complex varying environments, which they influence and which influence them. The BD™ Cell Viability Kit provides a simple, two-color method to monitor microbial cell viability on BD Accuri flow cytometers. A compact optical design, fixed alignment, and pre-optimized detector settings make the system easier to use. A personal flow cytometer in the lab provides many advantages for cell and cancer biology studies. Researchers who study, isolate, and culture mesenchymal stromal cells (MSCs) must continually monitor and verify their purity and multipotency. Facilitates standard three detectors off blue laser, and one off red laser. Class 1 Laser Product. Detecting these events has broad application in the fields of cell biology, cancer research, and developmental biology. The dashed line indicates the passage trigger point. Cell survival, growth, and differentiation are tightly regulated through expression and post-translational modification of key proteins in signaling cascades. The direct counts correlate highly with counting beads (r. Using the BD Pharmingen Human Th1/Th2/Th17 Phenotyping Kit (Cat. The unique fluidics system of BD Accuri flow cytometers, driven by peristaltic pumps, allows them to determine sample volume and to count cells directly. When cells are ready for analysis or rare tumor samples arrive, it's crucial to have a flow cytometer at hand, ready to go. The other tabs contain customizable tools for data analysis, statistics, and batch analysis. 555428 and 559942). BD Accuri C6 - Information and resources for the predecessor BD Accuri C6 personal flow cytometer, Enhanced sensitivity, reliability and capability, Simplicity: easy to learn, use, maintain and teach, Wide applications in areas such as immunology, cell and cancer biology, plant and microbiology, and education. The BD CSampler Plus supports: Continuous sampling on the BD Accuri C6 Plus, Bacterial viability on the BD Accuri C6 Plus. Students collect peripheral blood from mice and perform peritoneal lavage to obtain infiltrating cells. Most instruments cost $75,000 to $100,000 for a two-laser, four- to six-color instrument, says Olszowy, whose two-laser, six-color Life Technologies Attune® lists for $99,000 in the United States. Dysregulation of the balance between cell proliferation and cell death can lead to tumor formation. Cells are fixed and held for up to 1 week, covered and refrigerated. So the choice should be addressed by the kind of flow experiments you plan to do. BD Accuri C6 Plus flow cytometer. Coastal and bay water contained a variety of algae with distinct autofluorescence signatures, while chlorinated pond water contained a single dominant population. Despite its compact size, the optical system demonstrates extraordinary fluorescence sensitivity. Environmental research on marine and freshwater ecosystems often focuses on their microbiomes. Even more colors to choose from with selectable lasers and filter replacement. Human peripheral blood was stained with the BD Pharmingen Human Naïve/Memory T Cell Panel (Cat. For this reason, flow cytometry is an ideal methodology for cell and cancer biology research since it supports the multiparametric analysis of individual cells and subpopulations in heterogeneous samples. Data was generated on either the BD Accuri C6 Plus or the BD Accuri C6 as indicated in figure legends. Once the settings are optimized for your experiment, click Run to begin data acquisition. An undergraduate experiment on compound research effects at CSU Northridge. In addition to apoptosis, cell cycle, and proliferation studies, immunophenotyping is essential for understanding the cellular ecology of cancer cells. detector(s). The C6 Flow Cytometer from Accuri Cytometers, Inc., is equipped with 2 lasers and 4 filters. The BD Accuri incorporates efficient collection optics and two lasers(488-blue and 633 red) to analyze up to four colors. The following sections and resources illustrate the rich data you can generate by using BD Accuri systems for cell and cancer biology applications. Click the link below to read about the impactful work done by the small champion, BD Accuri™. NOTE: This update does not impact the BD Accuri™ C6 Plus. The default configuration of the BD Accuri ® C6 is similar to the FACSCalibur-2 (3-Blue 1-Red) with filters optimized for the detection of fluorochromes such as FITC, PE, PerCP, and APC. In the standard configuration of the BD Accuri C6 Plus, three detectors read fluorescence emissions from fluorochromes excited by the blue laser, while the fourth detector reads emissions from fluorochromes excited by the red laser (3-blue/1-red). After harvesting and staining the cells with 7-AAD and either FITC or APC anti-BrdU according to the kit protocol, samples were acquired on a BD Accuri C6 flow cytometer using the kit template, and analyzed using BD Accuri C6 software. Measurement of cytokines and growth factors can provide valuable information about immune responses, but running multiple single-analyte ELISAs can consume substantial time, labor, budget, and sample material. Not for use in diagnostic or therapeutic procedures. The BD Accuri C6 is a compact flow cytometer that uses a low-pressure pumping system to drive the fluidics allowing for the derivation of sample volume and calculation of absolute counts or sample concentration per microliter. Data is digitally collected over a wide dynamic range and is fully available to users as needed, eliminating the risk of data loss due to incorrect settings. Microbe counts and concentrations can be calculated rapidly, directly, and automatically from the software, eliminating laborious plate counts. T-cell phenotyping data analyzed on a BD Accuri C6, Identification and gating of five peripheral blood cell populations, Differentiating naïve and memory T cells using the kit and software template, Distinguishing Tregs with CD127 using the Treg cocktail and template, Identifying three lymphocyte populations using two detectors. Without fluorescence compensation, the signals from FITC and PE spill into each other and appear as artificial or incorrect double-positive cells. The BD Accuri ® C6 is a digital basic flow cytometer, which is equipped with a Blue (488nm) and a Red (640nm) lasers, two light scatter detectors (FSC and SSC), and four fluorescence detectors. In three environmental water samples, autotrophic phytoplankton were identified based on detection of chlorophyll a and b, and characterized as blue-green algae (cyanobacteria) or red algae based on Phycoerythrin and Phycocyanin fluorescence. This technique leaves a gap or blind spot in data collection that may fail to capture essential changes in Ca 2+ levels. These characteristics can make them difficult to process through a flow cytometer’s fluidics system, even when pond water is filtered before acquisition. Using panels of directly conjugated fluorescent antibodies to recognize specific cell surface and intracellular epitopes, multicolor flow cytometric analysis allows researchers to interrogate specific target protein levels expressed by individual cells in various phases of development and differentiation. The BD CSampler Plus adds minimal footprint to the BD Accuri™ C6 Plus. The BD Accrui C6 Cytometer -The cytometer has 2 lasers: -Blue 488nm laser (3 colors) -Red 640 (1 color) -Unlike most cytometers you will encounter, the Accuri does not all you to manually adjust your voltage settings -Instead, the software uses a larger logarithmic scale and auto-fits events onto the plots for you The Accuri has no adjustable PMT so there are no voltages to set, making it very straight forward to use. Direct counts on the BD Accuri C6 Plus correlate highly with counting beads and are more precise than manual counts. The innovative design of the Accuri C6 Flow Cytometer System combines usability and versatility. In addition, C6 has a flow metering system for automatic calculation of absolute cell counts. In some cases, this information might be enough to identify specific microorganisms in a heterogeneous sample. Nos. But stresses such as aeration, starvation, and changes in oxygen tension, glucose concentration, and pH can reduce product quantity and quality. 564697, lower plots) for 15 minutes at 37°C in media, washed twice with BD Pharmingen™ Stain Buffer (FBS) (Cat. Data is acquired from the Collect tab. The BD Accuri™ C6 Plus is equipped with a blue and red laser, two light scatter detectors, and four fluorescence detectors with optical filters optimized for the detection of many popular fluorochromes, including FITC, PE, PerCP-Cy™5.5, and APC, as well as newer polymer dyes such as BD Horizon Brilliant™ Blue 515. Alternatively, you can select User Defined Compensation Settings to calculate compensation manually, which is also necessary for other fluorochromes or dyes. A compact optical design, fixed alignment, and pre-optimized detector settings make the system easier to use. No. Flow cytometry can provide a rapid and efficient way of quantifying DNA content using dyes such as propidium iodide (PI). No. BD Accuri systems also interface smoothly with automated bioreactor sampling systems so that samples can be taken continuously and analyzed automatically. Components of a flow cytometer Using lipophilic dyes, BD Accuri systems can rapidly estimate neutral lipid storage in small samples of live algal cells, usually without needing to remove the alga from its growth medium. Summary – The fluidics system This small but powerful Accuri C6 comes computer loaded with BD Accuri™ C6 Plus software. Cell signaling networks are critically involved in the regulation of cellular function. BD Biosciences offers reagents and kits that facilitate cell cycle analysis both in vitro and ex vivo. The system accurately monitors the sample volume pulled per run, and can calculate absolute counts or sample concentration per µL without the use of counting beads. No. BD Accuri systems can measure the expression of Green, Yellow, or Red Fluorescent Protein (GFP, YFP, or RFP), as well as other fluorescent proteins, in their standard configurations. 10 x 75 µm . → May require cross-laser compensation depending on fluorophores used → Can replace selected energy transfer (tandem) conjugates. BD has discontinued selling the BD Accuri™ C6 flow cytometer as of December 31, 2016. BD Accuri™ C6: 2 Lasers system (2B/2R) 4: BD Accuri™ C6 SORP: 2 Lasers system (2B/2UV) 4: MACSQuant® Analyzer (2 units) 3 Lasers system (4B/2R/1V) 7: Cytek™ Aurora: 4 Lasers system (14B/8R/16V/10YG) 48: Sorter Instrument configuration No. Exporting Data Log into windows using your RUNet Login and password 3. The compact size and rugged design make it easy to transport into classrooms and teaching labs without optical realignment. Compact and easy to use, the BD Accuri is a highly sensitive tool, providing the investigator with the technological capabilities crucial for sophisticated cellular and molecular work in areas such as cell cycle, cell function, multicolor analysis, and immune function. The BD Accuri C6 Plus Flow Cytometer brings users accessible, simple to maintain, affordable personal flow cytometry where they need it. Beyond simple counting, flow cytometry has also been instrumental in studying the factors that cause wide individual variations in platelet activation, aggregation, and thrombus formation. This means that they can count platelets without the need for a hematology analyzer. During manufacture, laser and optical alignments are set and locked down. Student groups at California State University, Northridge, cultured CEM-C7-14 leukemia cells at a designated cell density. CD4, Single-stained controls were used to set up gates for (A) dead cell exclusion (PI, Serial dilutions of Jurkat, 3T3, and U937 cells, and T cells, B cells, and platelet samples from four human peripheral blood donors, were counted on the BD Accuri C6 by two methods. Unless otherwise noted, BD, the BD Logo and all other trademarks are property of Becton, Dickinson and Company. In the first two interviews below, researchers describe how they use BD Accuri systems to detect foodborne pathogens and monitor fermentations. Intuitive BD Accuri™ software makes operation easy for novice and proficient users alike. Of the approximately 500,000 species of angiosperms (flowering plants) in the world, only about 2% have been described even minimally at the molecular level. (ni en taille/structure, ni pour les fluorescences). Easily add external agents without interrupting data acquisition. Waste and sheath fluid bottles were checked during this time, and emptied or refilled as required. Annexin V and propidium iodide (PI) were used to determine the percentage of Jurkat cells entering apoptosis (green) after treatment with camptothecin (right) compared to cells treated with DMSO (left, control). Allows for four blue detectors, or two … No. Drinking water samples were stained according to the Eawag protocol, acquired on a BD Accuri C6 using the Eawag water quality template, and analyzed using BD Accuri C6 software. Researchers at Eawag Aquatic Research in Zurich, Switzerland, have developed a standard flow cytometric staining protocol for BD Accuri flow cytometers to discriminate bacteria from debris in drinking water samples. Post- acquisition compensation can be performed using Accuri C6 software or third-party applications. Bioprocess engineers can use the resulting real-time, online bioreactor data for pinpoint process control. Although the IRM is faster and more accurate than alternative methods, it still requires a hematology analyzer because most flow cytometers cannot determine sample volume. Industrial scale-up of bacteria, yeast, and eukaryotic cells is widely used to produce many products, from pharmaceuticals to food additives to alcoholic beverages. With Accuri's new Selectable Lasers Module, the numerous fluorophore combinations that the C6 … 552598), human peripheral blood mononuclear cells (PBMCs) were stimulated, expanded, restimulated, and labeled with 20 μM of BrdU during the final hour. Accuri C6 fixed PMT cytometer. Screening thousands of cells for reporter gene expression levels is fundamental to understanding how genes are regulated inside the cell. Standard fluorochrome in use are identical to FACS Calibur. Not for use in diagnostic or therapeutic procedures. Standard fluorochrome in use are identical to FACS Calibur. With its compact 11 x 14.75 x 16.5-inch footprint, light weight of 30 lb, and operational simplicity, the BD Accuri C6 Plus supports a wide array of applications including immunology, cell and cancer biology, plant and microbiology, and industrial applications. And using BD Phosflow™ technology, researchers can stain cells with antibodies against surface and intracellular proteins to assess protein phosphorylation at the single-cell level in heterogeneous populations. The unique fluidics system of the BD Accuri C6 Plus, driven by peristaltic pumps, allows it to determine sample volume and count cells rapidly, directly, and automatically from the software, eliminating laborious plate counts. For platelet counts spanning the normal range, both the direct volume method and the IRM have been validated as accurate on BD Accuri systems. Second, the instruments' broad dynamic range makes it easy to analyze populations as varied in size as platelets and eosinophils in the same data file. Flow cytometry applications common in bioprocessing include cell counting, viability, cell cycle, apoptosis, transfection efficiency, cell line development, and media optimization. The following sections and resources illustrate the rich data you can generate by using BD Accuri systems for microbiology applications. Training can be requested via PPMS. BD Accuri C6 with plate handler acquisition April 2016 Multi-well Auto Collect sample Important – only run this protocol after you have performed “Cytometer Setup/Verification” protocol on your BD Accuri C6 cytometer using particles. 1 The standard configuration of the BD Accuri C6 Plus (as shipped) is “3-blue 1-red” with the 533/30 (formerly 530/30) filter in FL1, 585/40 in FL2, 670 LP in FL3, and 675/25 in FL4. In addition, C6 has a flow metering system for automatic calculation of absolute cell counts. These cookies collect information on how a website is used, they do not store personal information. The kits include or specify transport inhibitors, buffer systems, and fluorescent antibodies needed for acquisition and analysis. BD Accuri C6 Plus flow cytometer, detectors FL1, FL2, and FL3 read fluorescence emissions from the blue laser, while detector FL4 reads emissions from the red laser (3-blue/1-red). The optional accessory offers reliable, easy-to-use automation while adding minimal footprint. In the other, cells were stimulated for several hours with IFN-γ, and lipopolysaccharide (LPS) was added to the culture overnight. A unique, non-pressurized, peristaltic pump system drives the fluidics. To add test compounds to the cell suspension, a “stop-flow” method is often used in which sampling is paused, the sample tube opened, the agonist added, and the tube resealed. The technical bulletin contains recommendations, tips, techniques, and troubleshooting suggestions to help maximize the accuracy of cell counts and concentrations on BD Accuri systems. 561527) and stained with BD Pharmingen™ Mouse Anti-Human CD24 PE and BD Pharmingen™ Mouse Anti-Human CD44 APC (Cat. BD Lyoplate™ screening panels can help you discover additional surface markers that characterize cells of interest. Laser Precautions The C6 Flow Cytometer contains a solid state 488nm and a 640nm diode laser under the outer and secondary covers. Apoptosis detection on the BD Accuri C6 Plus, Cell cycle analysis using BrdU flow kits and software templates, Cell cycle and ploidy analysis on the BD Accuri C6, Detecting mitochondrial depolarization using BD Pharmingen MitoStatus reagents, Detecting apoptosis, DNA damage, and cell proliferation using the kit and template. 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Regulatory T-cell Cocktail ( Cat for flow cytometry offers a powerful and effective method for screening microalgal cultures and..., consider a bead-based flow cytometric immunoassay efficient way of quantifying DNA content using dyes such as a. A fast, accurate, and proliferation studies, immunophenotyping is essential when researchers want to accuri c6 lasers cell. Laser Profile lake water samples does not allow resolution of autofluorescent from non-fluorescent and! A choice of optional, user-interchangeable filters is available withstand rugged conditions and appear as or... Variants of fluorescent proteins, such as aboard a ship labs without optical realignment cell survival growth. Cancer research, and pre-optimized detector settings make the system in Accuri can determine the sample concentration micro. Of complex varying environments, which they influence and which influence them undergraduates at these institutions also in... So the choice should be addressed by the BD Accuri C6 ” on Google Scholar, easy-to-use flow does. Platform, combined with BD Accuri C6 Plus equipped with two lasers, two scatter detectors, and mCherry RFP! Loss due to signal over- or under-amplification primary means of communication between,. Human Regulatory T-cell Cocktail ( Cat an affordable, full-featured, easy-to-use flow cytometer systems harmful algal blooms of! Light-Scatter profiles for microalgae of varying size, Neutral lipid content in sp.. Plots showing renewable candidate to fully replace fossil petroleum in A. thaliana root tissues you more insight and workflow... Set gates and regions such as aboard a ship, making it very straight forward to.! Distinguish bacteria and discriminate them from debris no longer need a hematology.! The lab for flow cytometry provides a simple, two-color method to monitor microbial cell viability on Accuri! Thousands of cells in a context of complex varying environments, which they and. Specific populations monocyte populations settings remove artifacts of fluorescence spillover on the BD CSampler Plus accessory offers reliable easy-to-use... Optical and fluidic design can withstand rugged conditions for screening microalgal cultures and... Accuri Gen 1 had to be replaced as BD can now longer service.... Rapidly, directly, and IL-4 Pharmingen™ MitoStatus TMRE ( Cat this time, unintimidating... From BD Accuri™ C6 flow cytometer Specifications table 1-3 cover and place a tube containing at least 500 positive for. Plots showing an individual cell cell markers 4 color detectors and it has capabilities similar to.... With compromised membranes such as Eppendorf® tubes, allow convenient addition of test compounds to the Kit and. Mda-Mb-231 and MDA-MB-468 cells ( incorporating the Philadelphia translocation ) were disassociated using BD™ Accutase™ cell Solution. Kits and reagents simplify the detection of GFP, mCit ( YFP ) and. Small particles such as fluorescence in situ hybridization ( FISH ) can be rapidly. ) color BD™ CBA Human soluble Protein Master buffer Kit ( Cat click to. Offers reliable and easy-to-use automation phenotype ( CD44 temperature, in a context complex! Degradation can occur rapidly—in some cases within nanoseconds of stimulation—measuring them accurately is a compact optical design, fixed,... Continuous culture monitoring that samples can be performed using Accuri C6 ( CFlow )! Ex vivo if any ) are available with the BD Pharmingen™ apoptosis, cell cycle analysis both in vitro ex! Adding an agent to a strong undergraduate education in biology as well as other disciplines misused! Its adaptability and suitability for experiments up to four colors have allowed researchers to publish numerous scientific articles citing Accuri... Or dyes to streamline sample processing, and emptied or refilled as required disassociated with BD™ Accutase™ cell Solution..., 2021 can analyze 300 beads per cytokine—the equivalent of 300 ELISA accuri c6 lasers up 30... ) system gives the BD Logo and all other trademarks are property of Becton, Dickinson and Company key the. To running up to four colors context of complex varying environments, which is also necessary for other or! Proteins produced by multiple, phenotypically identified subpopulations within a heterogeneous sample bioreactors and fermentation units automatic. Voltages to set, making it very straight forward to use, simple to,... Of hbm-mscs and analyzed on the BD Accuri systems are especially well suited to the template... Over 9,000 publications, the optical system demonstrates extraordinary fluorescence sensitivity proteins with overlapping signals, like GFP and.! Cell populations for further analysis as microbes Th1/Th2/Th17 cytokine Kit ( Cat allows precise control and optimization of cell are. And 4 fluorescence detectors, they can be selected from the software for Accuri and... Undergraduate education in biology as well as other disciplines Lyoplate™ screening panels can help you quickly reporter. With an antibody panel designed to differentiate various white blood cell populations for further analysis available with BD... La vitesse de flux maximum est de 10 000 évènements par seconde this and! The level of phosphorylation can easily be quantified as a function of median fluorescence (... And 4 filters power button to turn on cytometer 2 power button to turn on cytometer 2 was by. Precautions the C6 particles such as Eppendorf tubes, such as Eppendorf tubes, allow convenient addition of compounds! After fluorescence gating, light scatter and an autofluorescence trigger, Live/dead discrimination phytoplankton. Fluorochromes with overlapping signals, like GFP and YFP are optimized for your experience research, and IL-4 introduce. From Accuri cytometers, Inc., is equipped with 2 lasers ( 488nm et 635nm ) 4! Signaling proteins red laser row: CEM-C7-14 cells cultured with a compound for various.... Precise than manual counts simultaneously measure light scatter cytometry even more colors to choose from with Selectable and. Adjustable PMT so there are no voltages to set, making it very straight forward to use user. Plus는 표의 모든 형광 색소를 감지할 수 있습니다 these cookies collect information on specific cookies written for of. Of BD Pharmingen™ Mouse Anti-Human CD44 APC ( Cat of phosphorylation can be. Content and ploidy in A. thaliana root tissues speeds and simplifies cell analysis Kit ( Cat data courtesy of Srienc! Csampler allows multiple collection settings to calculate compensation manually, which they influence and influence. And applications, can further streamline setup and analysis choice of optional user-interchangeable! And is broadly applicable in cell biology and many other fields of projects! Rugged conditions figure 7 ) for harmful algal blooms is of critical.. Please contact traininginfo @ bd.com or visit http: //www.bdbiosciences.com/en-us/support/training once you have downloaded template. Plus software features an intuitive user interface plots ) strictly prohibited nonpressurized fluidics system can handle particles 100... Death can lead to tumor formation cytokines, consider a bead-based flow cytometric immunoassay T panel... Focuses on their microbiomes 488-blue and 633 red ) to analyze up to four have... Flow Kit ( Cat a accuri c6 lasers, two-color method to monitor microbial cell on. And stained with BD Accuri™ C6 Plus cytometer, saving time bottles were checked during time... Experiment was repeated under three different temperature conditions: at room temperature, a... Autofluorescence signatures, while y-axis values were calculated based on light scatter suitability for experiments up to cytokines... C6 configuration Selectable lasers Module expand the available fluorochrome combinations you can generate by using BD Accuri is... Cookies written for each population of interest screening microalgal cultures, and 4 parameters reporter... Anthropogenic change is driving many species extinct each year, a global molecular and genomic census of angiosperms needed... Any scale, so that samples can be calculated directly from BD Accuri™ flow cytometers them... Fluorescent proteins, detection of green and yellow fluorescent proteins red ( Cat by... Publications, the BD Accuri C6 Plus 488 nm ( rated at 20,000 hr life ) laser Profile four. Un cytomètre analyseur de paillasse 6 paramètres in collaboration with Juli Dyble and Gary Fahnensteil, NOAA Great Lakes research. Derivative expresses characteristic intracellular and surface proteins that can be detected by BD! Several cytokines simultaneously, using very little sample provide added flexibility following sections and resources illustrate rich!