BD ACCURI C6 PLUS
Among the array of applications that facilitate immunology and stem cell studies, multicolor flow cytometry is preeminent. Immunophenotyping was one of the first applications of flow cytometry, and for over 20 years, BD Biosciences has actively supported groundbreaking research in the field with flow cytometry systems and reagents.
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. BD’s solutions for cell identification, transcription factor expression, and cytokine secretion and measurement reflect a commitment to high quality and consistency needed for advanced research.
The BD Accuri™ C6 personal flow cytometer brings immunology and stem cell studies to the benchtop. With two lasers, two scatter detectors, and four fluorescence detectors, it can handle most common assays with ease. Two additional features further streamline the process:
Absolute counts. Cell concentration can be calculated directly from BD Accuri™ C6 software statistics tables without the addition of counting beads.
Kits and templates. BD Biosciences offers a range of cost-effective immunology and stem cell kits ideally suited for the BD Accuri C6 that include needed reagents and buffers to assess multiple cell surface and intracellular phenotyping markers. Free BD Accuri C6 software templates simplify data collection and analysis.
The following sections and resources illustrate the rich data you can generate by using the BD Accuri C6 for immunology and stem cell applications.
Flow cytometry can rapidly and accurately immunophenotype many kinds of cells, including lymphoid tissue (thymus, spleen, and lymph nodes), digested solid tissues, and blood.
For example, one common task in human blood analysis is to identify and enumerate populations of platelets, lymphocytes, monocytes, granulocytes, and eosinophils. Flow cytometry is a quick and reliable methodology for identifying these populations simultaneously in a single sample, based on their immunophenotypes (see table).
Several features of BD Accuri flow cytometers make them ideal platforms for differential analysis of human peripheral blood samples. First, fixed-voltage detectors simplify data collection and reduce the potential for data loss due to signal over- or under-amplification. 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. Third, the easy-to-use software Zoom tool allows precise control when setting gates. Finally, direct volume measurement allows direct calculation of the cell concentration per µL for each population of interest.
- Brochure: BD Accuri™ C6 Plus Flow Cytometer
- Technical Bulletin: Identification of Human Peripheral Blood Cell Populations with the BD Accuri™ Flow Cytometer
- Technical Bulletin: Taking Advantage of Fluorescence Spillover to Analyze More Markers on Fewer Detectors
- Product Information Sheet: BD Naïve/Memory T-Cell Kits and Templates
- Product Information Sheet: BD Regulatory T-Cell Kits and Templates
- Webinar Replay: Simplify Immunophenotyping Using Kits & Templates on the BD Accuri™ C6
The unique fluidics system of BD Accuri flow cytometers, driven by peristaltic pumps, allows them to determine sample volume, as well as count cells, directly. This feature speeds and simplifies cell analysis by calculating cell concentrations (per unit sample volume) automatically. Accurate cell concentrations are essential in many research applications, including enumerating leucocytes, B cells, T cells, and platelets in human blood, measuring microorganism concentrations in purified water, and determining the viability of cultured cell lines. Direct counts on BD Accuri systems correlate highly with counting beads, and are more precise than hemocytometer counts.
You can download two BD Biosciences documents on cell counting from the list below. The technical bulletin contains recommendations, tips, techniques, and troubleshooting suggestions to help maximize the accuracy of cell counts and concentrations on BD Accuri systems. The white paper presents sample data and guidance for viable cell concentrations in cultured cell lines, immune cell concentrations in human peripheral blood, and platelet counts in whole, unlysed human blood.
- Technical Bulletin: A Guide to Absolute Counting Using the BD Accuri™ Flow Cytometer
- White Paper: Determining Cell Concentration by Direct Volume on the BD Accuri™ Flow Cytometer
- Webinar Slide Set: Cell and Platelet Counting Using the BD Accuri™ C6 Flow Cytometer
Cytokine Analysis & Bead-Based Immunoassays
Cytokines and growth factors are a primary means of communication between cells, driving cellular differentiation that develops and supports the immune system. 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. Moreover, ELISAs average their results across the entire sample, obscuring differential cytokine production by different subpopulations of cells.
BD T-cell cytokine kits and reagents simplify the detection of cytokines using intracellular flow cytometry on the BD Accuri™ C6 flow cytometer. Intracellular flow cytometry allows the rapid analysis of cytokines and other inflammatory mediators produced by multiple, phenotypically identified subpopulations within a heterogeneous sample. It can also determine whether cytokine production by an activated cell population is the result of a few cells producing large amounts of cytokine or a large cell population producing small quantities. Finally, it can measure several cytokines simultaneously for an individual cell.
The kits include or specify transport inhibitors, buffer systems, and fluorescent antibodies needed for acquisition and analysis. Free BD Accuri™ C6 software templates matched to each kit include predefined workspaces, markers, regions, gates, and parameter names for quick and easy setup and analysis.
When you need to analyze a broad panel of cytokines, consider a bead-based flow cytometric immunoassay. Like ELISA assays, BD™ Cytometric Bead Array (CBA) assays measure secreted proteins produced by entire cell populations. But BD CBA can analyze 300 beads per cytokine—the equivalent of 300 ELISA wells—for up to 30 cytokines simultaneously, using very little sample. In essence, BD CBA is equivalent to running up to 30 ELISAs at once using flow cytometry.
- Product Information Sheet: BD Intracellular T-Cell Kits and Templates
- Product Information Sheet: BD™ Cytometric Bead Array (CBA) Reagents
- Article: Multiplex Cytokine Assays with BD™ CBA
- Webinar Replay: BD™ CBA on the BD Accuri™ C6
With the BD Accuri™ C6 flow cytometer, research laboratories no longer need a hematology analyzer to count platelets quickly and accurately.
Until 2001, platelets were commonly counted manually using a hemocytometer or automated impedance counter, despite the serious limitations of these methods. In 2001, two professional hematology organizations introduced an International Reference Method (IRM) for platelet counting based on flow cytometry. 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.
The unique fluidics system of the BD Accuri C6 flow cytometer, driven by peristaltic pumps, allows it to determine sample volume and to count cells directly. This means that it can count platelets without the need for a hematology analyzer. For platelet counts spanning the normal range, both the direct volume method and the IRM have been validated as accurate on the BD Accuri C6.
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.
- White Paper: Platelet Counting with the BD Accuri™ C6 Flow Cytometer
- Webinar Slide Set: Cell and Platelet Counting Using the BD Accuri™ C6 Flow Cytometer
- Customer Interview: Paul Harrison on Platelet Counting
- Customer Interview: Christopher Jones on Platelet Dynamics
BD Biosciences offers an extensive selection of hundreds of antibodies to stem cell pluripotency and differentiation markers that can be combined in many ways to monitor the cells’ changing expression patterns. BD Lyoplate™ screening panels can help you discover additional surface markers that characterize cells of interest.
A primary challenge of stem cell research is the inherent heterogeneity of the cell cultures. Researchers studying embryonic stem cells (ESCs) must constantly monitor the pluripotency of their cultures, while those studying human induced pluripotent stem cells (hiPSCs) must assess reprogramming success as well as pluripotency. Researchers who study, isolate, and culture mesenchymal stromal cells (MSCs) must continually monitor and verify their purity and multipotency.
Multicolor flow cytometry is ideal for this purpose because of its ability to interrogate heterogeneous cell populations, analyze their subpopulations, and characterize multiple antigens simultaneously. Each type of stem cell or derivative expresses characteristic intracellular and surface proteins that can be used for identification. Because intracellular analysis requires permeabilization, surface marker analysis is essential when researchers want to isolate live cell populations for further analysis.
- Product Information Sheet: BD Pluripotent Stem Cell Kits and Templates
- Product Information Sheet: BD Induced Pluripotent Stem Cell Kit and Template
- Product Information Sheet: BD Human Mesenchymal Stromal Cell Kit and Template
- Webinar Replay: Identification and Quality Control of Stem Cells Using Flow Cytometry (registration required)
- Customer Interview: Karina Palomares on Making Cardiomyocytes from Stem Cells
All reagents and kits are compatible with both the BD Accuri C6 Plus and BD Accuri C6 flow cytometer systems. Platforms referred to as "BD Accuri" represent both the BD Accuri C6 Plus and BD Accuri C6. Data was generated on either the BD Accuri C6 Plus or the BD Accuri C6 as indicated in figure legends.