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Flow Cytometry | Flow Cytometry Reagents | Bio-Techne

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Cytometry </h1></div> </div> <div class="field_block:paragraph:h1_2_2_text_to_media:field_lg_sngl_frmtd_text_global"> <div><p>Flow cytometry is a commonly used laser-based technique that is employed to count, sort, and phenotype cells in a single-cell suspension. Bio-Techne offers a broad-spectrum of antibodies and additional reagents and tools to streamline your flow cytometry experiments.</p></div> </div> </div> </div> <div class="layout__region layout__region--second"> <div class="hero-content-wrap"> <div class="field_block:paragraph:h1_2_2_text_to_media:field_indv_media"> <div> <img src="https://resources.bio-techne.com/bio-techne-assets/images/research-areas/bt-flow-cytometry-hero-img.png" alt="Flow Cytometry Hero Image" fetchpriority="high" > </div> </div> </div> </div> </div> </div> </div> </div> </div> </section> </div><!-- end hero-banner --> </div> </header> </div> <div id="main" class="container-fluid container-wide"> <div class="row"> <div class="col-md-12"> <div> <div id="block-bio-techne-global-breadcrumbs" class="system_breadcrumb_block"> <nav role="navigation" aria-labelledby="system-breadcrumb"> <h2 id="system-breadcrumb" class="visually-hidden">Breadcrumb</h2> <ol> <li> <a href="/">Home</a> </li> <li> <a href="/applications">Applications</a> </li> <li> Flow Cytometry </li> </ol> </nav> </div> </div> </div> </div> <main> <section class="section"> <div> <div id="block-bio-techne-global-content" class="system_main_block"> <article> <div> <div> <div> <div class="paragraph paragraph--type--freeform mb-4 paragraph--view-mode--default"> <div><p><strong><a href="#antibodies">Antibodies for Flow</a> |&nbsp;<a href="#cellstaining">Cell Staining and Detection</a> &nbsp;|&nbsp; <a href="#protocols">Staining Protocols</a> |&nbsp;<a href="#tools">Tools for Experimental Design</a> &nbsp;|&nbsp;<a href="#cellselection">Cell Selection</a> &nbsp;|&nbsp; <a href="#keyresources">Key Resources</a>&nbsp;| <a href="#background">Background &amp; FAQs</a></strong></p></div> </div> </div> <div> <div class="paragraph paragraph--type--freeform mb-4 paragraph--view-mode--default"> <div><h2>&nbsp;</h2><h2><a name="newproducts"></a>Featured Flow Cytometry Products</h2></div> </div> </div> <div> <div class="paragraph paragraph--type--section paragraph--view-mode--default"> <div> <div> <div class="paragraph paragraph--type--card-paragraph paragraph--view-mode--default card-para-default"> <div class="row card_row"> <div class="col-md-4"> <div class="h-100"> <div class="bg-white border rounded flex-column d-flex h-100"> <a href="/search?search_type=products&amp;keywords=mfluor%20antibody" > <div class="teaser-card-image-crop mb-3"> <img src="https://resources.bio-techne.com/bio-techne-assets/images/products/mfluor-voilet-conjugated-antibodies-header-525x315.jpg" alt="mFluor Violet Conjugated Antibodies " loading="lazy" > </div> </a> <a class="card_title_link" href="/search?search_type=products&amp;keywords=mfluor%20antibody" > <h3 class="card_title_htag"> <div><h3>Increased Flexibility in Flow Panel Design</h3></div> </h3> </a> <div class="card_description flex-grow-1"> <div><p>Explore an expansive catalog of mFluor™ Violet Conjugated Antibodies for enhanced versatility in your experiments.</p></div> </div> <div class="card-bottom-link"> <div> <a href="/search?search_type=products&amp;keywords=mfluor%20antibody" > Explore Now </a> </div> </div> </div> </div> </div> <div class="col-md-4"> <div class="h-100"> <div class="bg-white border rounded flex-column d-flex h-100"> <a href="/instruments/single-cell-dispensers" > <div class="card-img-top text-center mb-3" style="background-repeat: no-repeat; background-color: #f5f5f5; height:200px; width: 100%;"> <div class="teaser-valign-container" style=""> <div class="valign"> <img src="https://resources.bio-techne.com/bio-techne-assets/images/research-areas/bt-pala-side-instrument-525x300.png" alt="Bio-Techne Cell Sorter and Single Cell Dispenser side-angle" loading="lazy" > </div> </div> </div> </a> <a class="card_title_link" href="/instruments/single-cell-dispensers" > <h3 class="card_title_htag"> <div><h3>Fast, Gentle &amp; Easy Single Cell Isolation</h3></div> </h3> </a> <div class="card_description flex-grow-1"> <div><p>The Pala Cell Sorter and Single Cell Dispenser makes single cell sorting fast and easy. See how this benchtop instrument combine microfluidics, flow cytometry and direct liquid dispensing to accomplish gentle cell sorting and dispensing in one simple step.</p></div> </div> <div class="card-bottom-link"> <div> <a href="/instruments/single-cell-dispensers" > Learn More </a> </div> </div> </div> </div> </div> <div class="col-md-4"> <div class="h-100"> <div class="bg-white border rounded flex-column d-flex h-100"> <a href="/reagents/proteins/fluorokines-fluorescent-labeled-proteins" > <div class="card-img-top text-center mb-3" style="background-repeat: no-repeat; background-color: #f5f5f5; height:200px; width: 100%;"> <div class="teaser-valign-container" style=""> <div class="valign"> <img src="https://resources.bio-techne.com/bio-techne-assets/images/research-areas/flurokine-teaser-card.png" alt="Fluorokine teaser card" loading="lazy" > </div> </div> </div> </a> <a class="card_title_link" href="/reagents/proteins/fluorokines-fluorescent-labeled-proteins" > <h3 class="card_title_htag"> <div><h3>Fluorokines<sup>™</sup>: Fluorescently Labeled Proteins</h3></div> </h3> </a> <div class="card_description flex-grow-1"> <div><p>Easily measure transduction efficiency of CAR T or CAR NK cell preparations or determine B cell specificity after COVID-19 infection or vaccination.</p></div> </div> <div class="card-bottom-link"> <div> <a href="/reagents/proteins/fluorokines-fluorescent-labeled-proteins" > Learn More </a> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> <div> <div class="paragraph paragraph--type--freeform mb-4 paragraph--view-mode--default"> <div><h2><a name="antibodies"></a>Flow Cytometry Antibodies</h2></div> </div> </div> <div> <div class="paragraph paragraph--type--section paragraph--view-mode--default mb-4"> <div> <div> <div class="paragraph paragraph--type--_-2-media-to-text mb-4 paragraph--view-mode--default"> <div class="container-fluid container-wide"> <div class="layout layout--twocol-section layout--twocol-section--50-50"> <div class="layout__region layout__region--first"> <div class="hero-content-wrap"> <div class="field_block:paragraph:2_2_media_to_text:field_indv_media"> <div> <img src="https://resources.bio-techne.com/bio-techne-assets/images/research-areas/coloredvials2.png" alt="4 rows of fluorescently conjugated antibodies in vials with white and blue labels, and lids of various colors." loading="lazy" > </div> </div> </div> </div> <div class="layout__region layout__region--second"> <div class="hero-content-wrap"> <div class="field_block:paragraph:2_2_media_to_text:field_lg_sngl_frmtd_text_global"> <div><p>Find our full selection of flow cytometry validated antibodies, including fluorescent conjugated primary antibodies, secondary antibodies and isotype controls. Explore our broad selection of fluorescently conjugated antibodies, including Alexa Fluor® 488, Alexa Fluor® 647, PE, PE-Cy7™, and more! Take the guesswork out of flow cytometry panel design with our multicolor flow kits and panels.</p></div> </div> <div class="field_block:paragraph:2_2_media_to_text:field_call_to_action_global"> <a href="/search?keywords=antibody&amp;applications=Flow%20Cytometry" class="btn btn-primary" type="link"> Shop Flow Cytometry Antibodies </a> </div> </div> </div> </div> </div> </div> </div> <div> <div class="paragraph paragraph--type--freeform mb-4 paragraph--view-mode--default"> <div><p>&nbsp;</p> <figure class="responsive-figure-table"><table class="overflow-auto table table-bordered"> <tbody> <tr> <td class="w-25"><a href="/search?keywords=antibody&amp;applications=Flow%20Cytometry&amp;category_attr=Primary%20Antibodies">Primary Antibodies</a> &nbsp;</td> <td class="w-25"><a href="/search?keywords=antibody&amp;category_attr=Secondary%20Antibodies&amp;applications=Flow%20Cytometry">Secondary Antibodies</a>&nbsp;</td> <td class="w-25"><a href="/search?keywords=antibody&amp;category_attr=Isotype%20Controls">Isotype Controls</a>&nbsp;</td> <td><a href="/reagents/antibodies/multicolor-flow-cytometry-panels">Multicolor Kits and Panels</a>&nbsp;</td> </tr> </tbody> </table></figure></div> </div> </div> </div> </div> </div> <div> <div class="paragraph paragraph--type--freeform mb-4 paragraph--view-mode--default"> <div><h3><a name="cellstaining"></a>Additional Products for Cell Staining and Detection</h3> <figure class="responsive-figure-table"><table class="table table-bordered"> <tbody> <tr> <td class="w-33"><a href="/search?search_type=products&amp;keywords=apoptosis+detection+kit">Apoptosis Kits</a></td> <td><a href="/reagents/antibodies/epitope-tag-antibodies">Epitope Tag Antibodies&nbsp;</a></td> <td class="w-33"><a href="/search?keywords=fluorescent%20dna%20stains">Fluorescent DNA Stains</a></td> </tr> <tr> <td><a href="/reagents/fluorescent-dyes-probes/fluorescent-dyes">Fluorescent Dyes and Probes</a></td> <td><a href="/search?keywords=janelia%20fluor%20dyes">Janelia Fluor<sup>®</sup> Dyes</a></td> <td>&nbsp;<a href="/reagents/antibodies/conjugated-primary-antibodies/lightning-link-antibody-labeling-kits">Lighting-Link Antibody Labeling Kits</a></td> </tr> <tr> <td><a href="/reagents/fluorescent-dyes-probes/mitobrilliant-dyes">MitoBrilliant™ Mitochondrial Dyes</a></td> <td><a href="/applications/flow-cytometry/flow-cytometry-quality-control-beads">Quality Control Beads</a></td> <td><a href="/search?keywords=flow%20cytometry&amp;category_attr=Staining%20Kits%20and%20Reagents">Staining Kits and Buffers</a></td> </tr> </tbody> </table></figure></div> </div> </div> <div> <div class="paragraph paragraph--type--section paragraph--view-mode--default mb-4"> <div> <div> <div class="paragraph paragraph--type--freeform mb-4 paragraph--view-mode--default"> <div><h3><a name="protocols"></a>Flow Cytometry Staining Protocols</h3></div> </div> </div> <div> <div class="paragraph paragraph--type--_-3-media-to-text paragraph--view-mode--default mb-4"> <div class="container-fluid container-wide"> <div class="layout layout--twocol-section layout--twocol-section--25-75"> <div class="layout__region layout__region--first"> <div class="hero-content-wrap"> <div class="field_block:paragraph:1_3_media_to_text:field_indv_media"> <div> <img src="https://resources.bio-techne.com/bio-techne-assets/images/icons/protocols-faqs-icon525x315.png" alt="Protocols FAQs Icon" loading="lazy" > </div> </div> </div> </div> <div class="layout__region layout__region--second"> <div class="hero-content-wrap"> <div class="field_block:paragraph:1_3_media_to_text:field_lg_sngl_frmtd_text_global"> <div><ul> <li><a href="https://www.rndsystems.com/resources/protocols/flow-cytometry-protocol-staining-intracellular-molecules-using-detergents?_ga=2.17132857.1260935385.1705310688-171347606.1701252931" target="_blank">Cell Surface Staining Protocol</a>&nbsp;(at R&amp;D Systems)</li> <li><a href="https://www.rndsystems.com/resources/protocols/flow-cytometry-protocol-staining-intracellular-molecules-using-alcohol?_ga=2.42878245.1260935385.1705310688-171347606.1701252931" target="_blank">Intracellular Staining Protocol (Alcohols)</a>&nbsp;(at R&amp;D Systems)</li> <li><a href="https://www.rndsystems.com/resources/protocols/flow-cytometry-protocol-staining-membrane-associated-proteins-suspended-cells?_ga=2.17124025.1260935385.1705310688-171347606.1701252931" target="_blank">Intracellular Staining Protocol (Detergents)</a>&nbsp;(at R&amp;D Systems)</li> <li><a href="/applications/flow-cytometry/flow-cytometry-troubleshooting-guide">Flow Cytometry Trouble Shooting Guide</a></li> </ul></div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> <div> <div class="paragraph paragraph--type--section paragraph--view-mode--default"> <div> <div> <div class="paragraph paragraph--type--freeform mb-4 paragraph--view-mode--default"> <div><h2><a name="tools"></a>Flow Cytometry Tools for Experimental Design</h2></div> </div> </div> <div> <div class="paragraph paragraph--type--card-paragraph paragraph--view-mode--default card-para-default"> <div class="row card_row"> <div class="col-md-6"> <div class="h-100"> <div class="bg-white border rounded flex-column d-flex h-100"> <a href="/resources/flow-cytometry-panel-builder" > <div class="teaser-card-image-crop mb-3"> <img src="https://resources.bio-techne.com/bio-techne-assets/images/research-areas/novus-panel-builder-teaser-card-2.png" alt="Rainbow-colored mesh design over dark blue background." loading="lazy" > </div> </a> <a class="card_title_link" href="/resources/flow-cytometry-panel-builder" > <h3 class="card_title_htag"> <div><h3>Flow Cytometry Panel Builder Tool</h3></div> </h3> </a> <div class="card_description flex-grow-1"> <div><p>Save time and reduce costly mistakes by quickly finding compatible reagents using the Panel Builder Tool.</p></div> </div> <div class="card-bottom-link"> <div> <a href="/resources/flow-cytometry-panel-builder" > Build Your Panel </a> </div> </div> </div> </div> </div> <div class="col-md-6"> <div class="h-100"> <div class="bg-white border rounded flex-column d-flex h-100"> <a href="https://www.rndsystems.com/resources/cell-markers" > <div class="teaser-card-image-crop mb-3"> <img loading="lazy" class="card-background-image card-img-top" src="https://resources.bio-techne.com/bio-techne-assets/images/research-areas/cell-markers-teaser-card-background.png" /> </div> <div class="card-foreground-image"> <img src="https://resources.bio-techne.com/bio-techne-assets/images/research-areas/cell-markers-teaser-card.png" alt="Cell Markers teaser card" loading="lazy" > </div> </a> <a class="card_title_link" href="https://www.rndsystems.com/resources/cell-markers" > <h3 class="card_title_htag"> <div><h3>Interactive Cell Marker Tool</h3></div> </h3> </a> <div class="card_description flex-grow-1"> <div><p>Learn more about cell surface and intracellular markers from R&amp;D Systems, a Bio‑Techne brand.</p></div> </div> <div class="card-bottom-link"> <div> <a href="https://www.rndsystems.com/resources/cell-markers" > Explore Interactive Cell Marker Tool at R&amp;D Systems </a> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> <div> <section class="paragraph paragraph--type--full-width-banner-with-image mb-4 paragraph--view-mode--default paragraph-background-color--"> <div class="banner_background_image" data-bgimage="https://resources.bio-techne.com/bio-techne-assets/images/banners/MitoBrillant-banner.jpg" style="background-repeat: no-repeat;"> <div> <div> <div> <div class="paragraph paragraph--type--_-2-text-to-media mb-4 paragraph--view-mode--default"> <div class="container-fluid container-wide"> <div class="layout layout--twocol-section layout--twocol-section--50-50"> <div class="layout__region layout__region--first"> <div class="hero-content-wrap"> <div 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</div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </section> </div> <div> <div class="paragraph paragraph--type--section paragraph--view-mode--default mb-4"> <div> <div> <div class="paragraph paragraph--type--freeform mb-4 paragraph--view-mode--default"> <div><h3>MagCellect™ Cell Selection Kits &amp; Reagents</h3> <p>Isolate and enrich cell populations through magnetic, bead-based positive or negative selection without the need for specialized columns.</p> <h4>Featured MagCellect Kits</h4> <figure class="responsive-figure-table"><table class="table table-bordered"> <tbody> <tr> <td><a href="/t/b-cell-isolation-kits/cell-selection-detection-kits">B Cells</a></td> <td><a href="/search?keywords=magcellect&amp;common_name=T%20Cell%20Isolation%20Kits|CD3|CD4">CD4+ T Cells</a></td> <td><a href="/t/cd45/cell-selection-detection-kits">CD45+ Cells</a></td> <td><a href="/t/natural-killer-nk-cell-isolation-kits">NK Cells</a></td> </tr> <tr> <td><a href="/search?keywords=magcellect&amp;common_name=T%20Cell%20Isolation%20Kits|CD3">CD3+ T Cells</a></td> <td><a href="/search?keywords=magcellect&amp;common_name=CD25/IL-2R%20alpha">CD4+ CD25+ Tregs</a></td> <td><a href="/search?keywords=magcellect&amp;common_name=CD44">CD44+ Cells</a></td> <td><a href="/p/cell-selection-detection-kits/magcellect-plus-human-egfr-cell-isolation-kit_magh124">EGFR+ Cells</a></td> </tr> <tr> <td><a href="/t/cd8/cell-selection-detection-kits">CD8+ T Cells</a></td> <td><a href="/p/cell-selection-detection-kits/magcellect-human-cd14-cell-isolation-kit_magh105">CD14+ Monocytes</a></td> <td><a href="/p/cell-selection-detection-kits/magcellect-plus-human-epcam-cell-isolation-kit_magh121">EpCAM+ Cells</a></td> <td><a href="/p/cell-selection-detection-kits/magcellect-plus-human-e-cadherin-cell-isolation-kit_magh123">E-Cadherin+ Cells</a></td> </tr> </tbody> </table></figure></div> </div> </div> <div> <div class="paragraph paragraph--type--individual-button mb-4 paragraph--view-mode--default w-100"> <div class="row"> <div class="col"> <a href="/search?keywords=magcellect" class="btn btn-primary float-left" type="link" > Browse all MagCellect Kits </a> </div> </div> </div> </div> </div> </div> </div> <div> <div class="paragraph paragraph--type--freeform mb-4 paragraph--view-mode--default"> <div><h2><a name="keyresources"></a></h2> <h2>Key Flow Cytometry Resources</h2></div> </div> </div> <div> <div class="paragraph paragraph--type--section paragraph--view-mode--default"> <div> <div> <div class="paragraph paragraph--type--_-3-media-to-text paragraph--view-mode--default mb-4 p-3"> <div class="container-fluid container-wide"> <div class="layout layout--twocol-section layout--twocol-section--25-75"> <div class="layout__region layout__region--first"> <div class="hero-content-wrap"> <div class="field_block:paragraph:1_3_media_to_text:field_indv_media"> <div> <img src="https://resources.bio-techne.com/bio-techne-assets/images/research-areas/image-flow-cytometry-handbook_480x485_Feb2023Update.png" alt="View of the Flow cytometry handbook and a single cell as a background" loading="lazy" > </div> </div> </div> </div> <div class="layout__region layout__region--second"> <div class="hero-content-wrap"> <div class="field_block:paragraph:1_3_media_to_text:field_header_2_global"> <div><h3>Flow Cytometry Handbook</h3></div> </div> <div class="field_block:paragraph:1_3_media_to_text:field_lg_sngl_frmtd_text_global"> <div><p>Our Flow Cytometry Handbook is the must have resource for flow cytometry researchers. Download now to find step-by-step protocols and troubleshooting tips.</p></div> </div> <div class="field_block:paragraph:1_3_media_to_text:field_call_to_action_global"> <a href="https://info.bio-techne.com/flow-cytometry-handbook.html" class="btn btn-primary" type="link"> Download Now </a> </div> </div> </div> </div> </div> </div> </div> <div> <div class="paragraph paragraph--type--_-3-media-to-text paragraph--view-mode--default mb-4 p-3"> <div class="container-fluid container-wide"> <div class="layout layout--twocol-section layout--twocol-section--25-75"> <div class="layout__region layout__region--first"> <div class="hero-content-wrap"> <div class="field_block:paragraph:1_3_media_to_text:field_indv_media"> <div> <img src="https://resources.bio-techne.com/bio-techne-assets/images/icons/videos-webinars-icon525x315.png" alt="Videos Webinars Icon" loading="lazy" > </div> </div> </div> </div> <div class="layout__region layout__region--second"> <div class="hero-content-wrap"> <div class="field_block:paragraph:1_3_media_to_text:field_header_2_global"> <div><h3> Webinars</h3></div> </div> <div class="field_block:paragraph:1_3_media_to_text:field_lg_sngl_frmtd_text_global"> <div><ul> <li><a href="https://www.rndsystems.com/blog/relax-and-go-flow-demystifying-multi-parameter-flow-cytometry?_ga=2.8743221.1260935385.1705310688-171347606.1701252931">Flow Cytometry Webinar Part 1</a></li> <li><a href="https://www.rndsystems.com/blog/webinar-2-turning-flow-cytometry-upside-down-and-inside-out-cellular-function-revealed?_ga=2.13479479.1260935385.1705310688-171347606.1701252931">Flow Cytometry Webinar Part&nbsp;2</a></li> </ul></div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </div> <div> <div class="paragraph paragraph--type--freeform mb-4 paragraph--view-mode--default"> <div><h2><a name="background"></a></h2> <h2>Background Information, FAQs, Resources and Products</h2></div> </div> </div> <div> <div class="paragraph paragraph--type--section paragraph--view-mode--default mb-4"> <div> <div> <div class="paragraph accordion mb-4 paragraph--type--extendable-paragraph paragraph--view-mode--default"> <div class="card-header rounded-top bg-white"> <a class="collapsed" data-toggle="collapse" href="#expPara-about-flow-cytometry8" role="button" aria-expanded="false" aria-controls="expPara"> <h5>About Flow Cytometry</h5> </a> </div> <div class="collapse" id="expPara-about-flow-cytometry8" aria-expanded="false" aria-controls="expPara"> <div class="card-body p-4"> <br/> <div><p>Flow cytometry is a high throughput, single-cell analysis technique that uses lasers to rapidly&nbsp;activate fluorescently-labeled conjugated antibodies on cells in a suspended solution. It is a powerful tool as it can measure many parameters on single cells, as well as very quickly collect information from millions of cells. In flow cytometry, single-cell suspensions are passed through single or multiple lasers, which causes the light to scatter. Cell populations can be sorted based on light scatter, where forward scatter provides cell size analysis and side scatter determines cell granularity. Flow cytometry can provide information about cell size and volume, evaluate the purity of isolated cell subpopulations, characterize different cell types in a heterogeneous population, while providing a technique that can examine the expression of both cell surface and intracellular proteins.</p> <p>Fluorophore compounds, such as fluorescently-labeled antibodies or propidium iodide, are commonly used in flow cytometry to detect proteins of interest. Using fluorophores with nonoverlapping maximum emission spectra enables simultaneous individual detection of&nbsp; fluorescently stained cells. Isotype control antibodies are used as negative controls to validate a flow cytometry experiment. Multiple research areas including cancer biology, immunology, infectious disease monitoring, virology, and molecular biology utilize flow cytometry.</p></div> </div> </div> </div> </div> <div> <div class="paragraph accordion mb-4 paragraph--type--extendable-paragraph paragraph--view-mode--default"> <div class="card-header rounded-top bg-white"> <a class="collapsed" data-toggle="collapse" href="#expPara-faqs7" role="button" aria-expanded="false" aria-controls="expPara"> <h5>FAQs</h5> </a> </div> <div class="collapse" id="expPara-faqs7" aria-expanded="false" aria-controls="expPara"> <div class="card-body p-4"> <br/> <div><p><strong>What is FACS?</strong></p> <p>Fluorescence-activated cell sorting (FACS) is a specialized type of flow cytometry. It provides a method for sorting a heterogeneous mixture of biological cells into separate containers, one cell at a time. Cell sorting is based upon the specific light scattering and fluorescent characteristics of each cell.</p> <p><strong>What is the minimum number of cells needed for flow cytometry?</strong></p> <p>The number of cells needed for a particular experiment is dependent on your needs. You would need fewer cells if you are probing for a robustly expressed antigen compared to a sparsely expressed protein. Additionally, you would need more cells for multicolor flow cytometry versus single color staining. However, we recommend starting with 1 × 10<sup>6</sup>&nbsp;cells per 100 µl of staining buffer per tube.</p> <p><strong>What controls are used in flow cytometry?</strong></p> <p>Flow cytometry controls can be split into the following categories.</p> <ol> <li>&nbsp;<strong>Instrument Controls:</strong> Calibration particles or fluorescent beads from the instrument’s manufacturer should be run prior to use to ensure the instrument is performing properly.</li> <li><strong>Compensation Controls</strong>: When performing multicolor flow cytometry, the emission spectra from the different fluorophores used can overlap and can spread into multiple channels. The process of correcting for spillover is known as compensation. For compensation, a single stained sample is needed from each fluorochrome in the experiment. These can be set up using cells, compensation beads, or antibody capture beads.</li> <li><strong>Gating Controls</strong>: One of the most critical aspects of analyzing flow cytometry data is properly setting gates. The Fluorescence Minus One (FMO) control is necessary for interpreting flow data and for setting gates. Since FMO controls provide a reliable assessment of background in a multicolor experiment, they can accurately discriminate between negative and positive cell populations.</li> <li><strong>Cell Viability Controls</strong>: Dead cells can produce false positives because they autofluoresce and display an increased affinity for non-specific antibody binding. As a result, these dead cells need to be removed from data analysis. Several fluorescent dyes, such as 7-AAD, calcein AM, and propidium iodide, can be used to distinguish dead cells from live cells.</li> <li><strong>Unstained Cells Controls:</strong>&nbsp;Several cell components, such as flavin coenzymes and NADPH, are endogenous fluorophores that exhibit autofluorescence. Running a sample of unstained cells through the flow cytometry will allow you to determine the amount of background fluorescence due to autofluorescence so that you can gate appropriately.</li> <li><strong>Isotype Controls:</strong> Isotype controls are used to assess non-specific binding due to antibody class. An isotype control is an antibody that lacks specificity to the target of interest, but matches the class, type, and fluorescent conjugate of the primary antibody.</li> </ol></div> </div> </div> </div> </div> <div> <div class="paragraph accordion mb-4 paragraph--type--extendable-paragraph paragraph--view-mode--default"> <div class="card-header rounded-top bg-white"> <a class="collapsed" data-toggle="collapse" href="#expPara-additional-products-and-resources-for-flow-cytometry-workflow24" role="button" aria-expanded="false" aria-controls="expPara"> <h5>Additional Products and Resources for Flow Cytometry Workflow</h5> </a> </div> <div class="collapse" id="expPara-additional-products-and-resources-for-flow-cytometry-workflow24" aria-expanded="false" aria-controls="expPara"> <div class="card-body p-4"> <br/> <div><p><strong>Products for Differentiating and Expanding Cells</strong></p> <ul> <li><a href="/reagents/cell-culture-reagents">Cell Culture Reagents</a></li> <li><a href="/search?keywords=supplement&amp;category_attr=Cell%20Culture%20Products">Cell Culture Media Supplements</a></li> <li><a href="/search?keywords=cellxvivo">CellXVivo Differentiation and Expansion Kits</a></li> </ul> <p><strong>Resources</strong></p> <ul> <li><a href="/reagents/antibodies/immune-cell-therapy-antibodies">Antibodies for Immune Cell Therapy</a></li> <li><a href="https://resources.rndsystems.com/images/site/rnd-systems-bcells-br.pdf?_ga=2.244881093.1317747681.1643637958-1474006042.1618344313&amp;_gac=1.116232692.1642705714.Cj0KCQiAraSPBhDuARIsAM3Js4oG-Ir_IonY5fPN7uKauC1VKMPCKfoJJMuYUP09T6yfYcksOfxMLCkaAkjNEALw_wcB" target="_blank">B Cells Brochure</a></li> <li><a href="https://resources.rndsystems.com/images/site/rnd-systems-cd4-br.pdf?_ga=2.244881093.1317747681.1643637958-1474006042.1618344313&amp;_gac=1.116232692.1642705714.Cj0KCQiAraSPBhDuARIsAM3Js4oG-Ir_IonY5fPN7uKauC1VKMPCKfoJJMuYUP09T6yfYcksOfxMLCkaAkjNEALw_wcB" target="_blank">CD4+ T cell subsets Brochure</a></li> <li><a href="/resources/literature/human-immune-cell-marker-poster">Cell Markers Guide for Human Immune Cell Characterization</a></li> <li><a href="/reagents/antibodies/conjugated-primary-antibodies/tandem-dye-conjugated-antibodies-faqs">Tandem Dye Conjugated Antibodies FAQs</a></li> </ul></div> </div> </div> </div> </div> </div> </div> </div> </div> </div> </article> </div> </div> </section> </main> </div> <div id="brandsbar-footer" class="bg-bt-primary"> <div class="container-wide container-fluid text-white"> <div class="row py-5"> <div class="footer-logo col-xl-3 col-lg-auto d-flex"> <img class="brand-bar-footer-logo" src="/themes/custom/bio_techne_global/logo.svg" alt="Bio-techne logo"> </div> <div class="col-xl-9 col-lg-auto"> <p class="ml-xl-4 bt-px_groteskbold footer-tagline">Global Developer, Manufacturer, and Supplier of High-Quality Reagents, Analytical Instruments, and Precision Diagnostics.</p> </div> <div class="included-brands col"> <span class="font-weight-bold mr-3 bt-inter_regular">INCLUDES</span> <span class="mr-3 bt-inter_regular">R&D Systems™</span> <span class="mr-3 bt-inter_regular">Novus Biologicals™</span> <span class="mr-3 bt-inter_regular">Tocris Bioscience™</span> <span class="mr-3 bt-inter_regular">ProteinSimple™</span> <span class="mr-3 bt-inter_regular">ACD™</span> <span class="mr-3 bt-inter_regular">ExosomeDx™</span> <span class="mr-3 bt-inter_regular">Asuragen™</span> <span class="mr-3 bt-inter_regular">Lunaphore™</span> </div> </div> </div> </div> <div id="site-footer__top" class=" pt-4"> <div class="container-wide container-fluid pt-3"> <div> <div class="col-md-12"> <footer> <div> <div class="row"> <div class="col-xl-3 col-md-6 col-xs-12 p-0 text-white bt-inter_regular"> <div> <div id="block-footercolumn1" class="block_content:8da4adc8-094f-4085-8706-613703de4da6"> <div><h4>CONTACT INFORMATION</h4> <p>614 McKinley Place NE<br /> Minneapolis, MN 55413<br /> USA</p> <p>TEL <a href="tel:612 379 2956">612 379 2956</a><br /> Toll-free <a href="tel:800 343 7475">800 343 7475</a><br /> FAX 612 656 4400</p> </div> </div> </div> </div> <div class="col-xl-3 col-md-6 col-xs-12 p-0 bt-inter_regular"> <div> <nav role="navigation" aria-labelledby="block-footercolumn2-menu" id="block-footercolumn2"> <h4 id="block-footercolumn2-menu">Who We Are</h4> <ul> <li> <a href="/about/about-bio-techne" title="About Bio-Techne" data-drupal-link-system-path="node/16">About Bio-Techne</a> </li> <li> <a href="/brands" title="Brands" data-drupal-link-system-path="node/46">Bio-Techne Brands</a> </li> <li> <a href="/about/careers" title="History" data-drupal-link-system-path="node/21">Careers </a> </li> <li> <a href="/about/corporate-and-social-responsibility" title="Corporate Responsibility" data-drupal-link-system-path="node/61">Corporate &amp; 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