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RNAscopePlusLSAssay | In Situ Hybridization, RNA-ISH | ACDBio

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src="https://acdbio.com/sites/default/files/STRY0320271_SBD_WU_ACDbio-RNAscope_Automated_Assays_TA-min.jpg" class="img-banner"><h1 class="general-title"></h1><h2 class="banner-sub-title-seo"></h2></div><div class="content-container container"><div class="region region-content"><div id="block-system-main" class="block block-system clearfix"><div class="content"><div id="node-464323" class="node node-product-sub-category node-promoted page-node- node-type-product-category" about="/rnascopepluslsassay" typeof="sioc:Item foaf:Document"><div class="nav-tab-container"><ul class="nav nav-tabs page-tabs" id="myTab"><li><a href="/automated-assay-leica">Overview</a></li><li class="pro-category-20224" ><a href="/rnascope-25-ls-assay-brown">RNAscope 2.5 LS Assay-BROWN</a></li><li class="pro-category-20225" ><a href="/rnascope-25-ls-assay-red">RNAscope 2.5 LS Assay-RED</a></li><li class="pro-category-104792" ><a href="/rnascope-25-ls-duplex-assay">RNAscope 2.5 LS Duplex Assay</a></li><li 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class="profile-content"><h3>RNAscope™ Plus smRNA-RNA LS Assay</h3></div><div class="profile-content"><div class="field field-name-body field-type-text-with-summary field-label-hidden"><div class="field-items"><div class="field-item even" property="content:encoded"><p>Small regulatory RNAs, including miRNAs, are instrumental in regulating human health and have been associated with a wide range of different diseases. Oligonucleotide therapies, including antisense oligonucleotides (ASOs) and RNA interference (RNAi), represent a unique class of gene therapies that have the potential to provide highly focused, long-lasting therapeutic effects by utilizing similar cellular processes.</p><p>Advanced Cell Diagnostics miRNAscope ISH technology has become the gold standard for identifying the cellular localization of endogenous miRNAs and measuring delivery efficiency of therapeutic oligonucleotides in preclinical animal models. Combining proprietary RNAscope and miRNAscope ISH technologies, we are now introducing the RNAscope Plus smRNA-RNA LS assay which is designed as an automated assay on the Leica Biosystems' BOND RX Research Advanced Staining System.  It is a valuable new tool to quantify changes in gene expression and cellular function in response to the introduction of regulatory RNAs, essential for optimization of therapeutic efficacy and safety.</p><p> </p><p><strong>Key Features:</strong></p><ul><li>Unlock the potential to detect 1 small RNA along with up to 3 mRNA targets on the Leica Biosystems' BOND RX automated staining system</li><li>Visualize antisense oligonucleotides (ASOs), microRNAs (miRNAs), small interfering RNAs (siRNAs), and other smaller RNAs that are 17-50 nt in length along with up to 3 mRNA targets</li><li>Visualize gene regulation with morphological context</li><li>Validate miRNA biomarkers in intact tissues</li><li>Assess small RNA therapeutic delivery and biodistribution</li><li>Add a visual dimension to interrogate heterogeneous tissues and evaluate therapeutic efficacy.</li><li>Combined ACD patented signal amplification system and the Tyramide Signal Amplification (TSA™) technology</li></ul><p> </p><div class="container overflow-hidden"><div class="row review app-note gx-3 flyer-brochure"><div class="col-sm-6"><h3>Brochure</h3><div class="col-md-5 p-0"><img src="/sites/default/files/Introducing-RNAscope-Plus-small-RNA-RNA-Fluorescent-Assay.jpg" style="max-width: 100%;" /></div><div class="col-md-7 download-pdf"><p> </p><h4>Introducing the RNAscope Plus small RNA-RNA Fluorescent Assay</h4><p><a class="down-pdf-btn learn-more" href="/system/files_force/STRY0300808_SBD_BR_RNAscope-Plus_TA.pdf?download=1">Download Brochure</a></p></div><div class="clear"> </div></div><div class="col-sm-6"><h3>Flyer</h3><div class="col-md-5 p-0"><img src="/sites/default/files/STRY0292871_SBD_FL_RNAscope-Plus.jpg" style="max-width: 100%;" /></div><div class="col-md-7 download-pdf"><h4>Biodistribution, safety and efficacy assessment of small RNAs with the new RNAscopeTM Plus small RNA-RNA co-detection Assay</h4><p><a class="down-pdf-btn learn-more" href="/system/files_force/STRY0292871_SBD_FL_RNAscope-Plus.pdf?download=1">Download Flyer</a></p></div><div class="clear"> </div></div></div></div><p> </p><p><strong>Technology Overview:</strong></p><p>RNAscope Plus smRNA-RNA LS assay allows simultaneous detection of 1 smRNA and up to 3 RNA targets within a single sample using up to four target probes and the nuclei can be counterstained and visualized using DAPI fluorescent dye. The four target probes, each designed in a different probe channel—S1 for smRNA, and C2, C3, or C4 for RNA targets. The assay is compatible with formalin-fixed, paraffin-embedded (FFPE) and fresh frozen (FF) tissue.</p><p>The key technological advantage of this assay is the <strong>significant signal boost with simultaneous background noise suppression</strong> which is due to advances in ACD’s patented signal amplification technology combined with Tyramide Signal Amplification (TSA™) technology.</p><p> </p><p><strong>Guidelines for Microscope Channel Selection:</strong></p><p>RNAscope Plus smRNA-RNA LS assay supports interrogation of up to 4 target probes. The assay uses four target probes with each probe designed in a different channel – the S1 channel for smRNA and C2, C3 and/or C4 channels for other RNA targets. The nuclei can be counterstained and visualized using DAPI fluorescent dye. For viewing of 4 plex assay, a multiplexed biomarker imaging system is required, such as Nuance FX Multiplex Biomarker Imaging System, Vectra 3.0 Automated Quantitative Pathology Imaging System or Mantra Quantitative Pathology Workstation Akoya Biosciences.</p><p>Fluorophore options depend on the imaging systems available. Two color combination examples are shown in the following table. Multispectral un-mixing capability are required for certain options.</p><table class="classification full"><thead><tr><th>Microscopy Channel</th><th>Enzyme</th><th>Fluorophores Option 1</th><th>Fluorophores Option 2</th></tr></thead><tbody><tr><td>S1</td><td>HRP</td><td>Opal™ 690nm</td><td>TSA Vivid 650nm</td></tr><tr><td>C2</td><td>HRP</td><td>Opal™  520nm</td><td>TSA Vivid 520nm</td></tr><tr><td>C3</td><td>HRP</td><td>Opal™  570nm</td><td>TSA Vivid 570nm</td></tr><tr><td>C4</td><td>HRP</td><td>Opal™  780nm</td><td>Opal™ 780nm</td></tr></tbody></table><p> </p><p><strong>What to order</strong><br />Visualization requires a multispectral fluorescent imaging system (e.g. Nuance FX Multiplex Biomarker Imaging System)</p><p> </p><p><strong>Order the following components from ACD:</strong></p><table class="classification full"><thead><tr><th scope="col">Component</th><th scope="col">Cat. No.</th></tr></thead><tbody><tr><td>RNAscope™ Plus smRNA-RNA LS Reagents Kit (Includes RNAscope™ Plus  smRNA-RNA LS Detection Kit and RNAscope LS Multiplex TSA Buffer Pack)</td><td>322786</td></tr><tr><td>miRNAscope LS Target Probes (Catalog or Made-to-Order LS-S1 Probes)</td><td>Various</td></tr><tr><td>RNAscope LS 2.5 Target Probes<br />(Catalog or Made-to-Order C2, C3 &amp; C4 LS 2.5 Probes)</td><td>Various</td></tr><tr><td>RNAscope Plus smRNA-RNA 4-plex LS Positive Control Probe-Hs</td><td>323368</td></tr><tr><td>RNAscope Plus smRNA-RNA 4-plex LS Positive Control Probe-Mm</td><td>323378</td></tr><tr><td>RNAscope Plus smRNA-RNA 4-plex LS Positive Control Probe-Rn</td><td>323388</td></tr><tr><td>RNAscope Plus smRNA-RNA 4-plex LS Negative Control Probe</td><td>323398</td></tr></tbody></table><p> </p><p><strong>Fluorophores required to perform RNAscope smRNA-RNA LS Fluorescent Assay</strong></p><table class="classification full"><thead><tr><th scope="col">Fluorophores </th><th scope="col">Part number (ACD)</th><th scope="col">Recommended dilution range</th></tr></thead><tbody><tr><td>TSA Vivid Fluorophore 520</td><td>323271 TSA Vivid Fluorophore 520 (ACD)</td><td>1:750–1:3000</td></tr><tr><td>TSA Vivid Fluorophore 570</td><td>323272 TSA Vivid Fluorophore 570</td><td>1:750–1:3000</td></tr><tr><td>TSA Vivid Fluorophore 650</td><td>323273 TSA Vivid Fluorophore 650</td><td>1:750–1:3000</td></tr></tbody></table><p> </p><table class="classification full"><thead><tr><th scope="col">Fluorophores </th><th scope="col">Part number (Akoya Bioscience)</th><th scope="col">Recommended dilution range</th></tr></thead><tbody><tr><td>Opal 520</td><td>FP1487001KT: Opal 520 Reagent Pack</td><td>1:750–1:3000</td></tr><tr><td>Opal 570</td><td>FP1488001KT: Opal 570 Reagent Pack</td><td>1:750–1:3000</td></tr><tr><td>Opal 690</td><td>FP1497001KT: Opal 690 Reagent Pack</td><td>1:750–1:3000</td></tr><tr><td>Opal Polaris 780</td><td>FP1501001KT: Opal Polaris 780 Reagent Pack </td><td>TSA-DIG#  : 1:750–1:3000<br />Polaris 780: 1:187.5–1:750 </td></tr></tbody></table><p> </p><p><strong>Required materials and equipment from Leica BOND RX</strong><br />The RNAscope smRNA-RNA LS Fluorescent Assay requires specific materials and equipment available only from Leica Biosystems.</p><table class="classification full"><thead><tr><th scope="col">Component</th><th scope="col">Cat. No.</th></tr></thead><tbody><tr><td>Leica Biosystems’ BOND RX System — automated slide stainer</td><td>—</td></tr><tr><td>BOND 30 mL Open containers </td><td>OP309700</td></tr><tr><td>BOND 7 mL Open containers* </td><td>OP79193</td></tr><tr><td>BOND Research Detection System</td><td>DS9455</td></tr><tr><td>BOND Universal Covertile</td><td>S21.4611</td></tr><tr><td>BOND Epitope Retrieval Solution 1-1L (RTU)</td><td>AR9961</td></tr><tr><td>BOND Epitope Retrieval Solution 2-1L (RTU)</td><td>AR9640</td></tr><tr><td>BOND Dewax Solution – 1L (RTU)</td><td>AR9222</td></tr><tr><td>BOND Wash Solution 10X Concentrate – 1L</td><td>AR9590</td></tr><tr><td>BOND Aspirating Probe Cleaning System</td><td>CS9100</td></tr><tr><td>BOND Mixing Stations</td><td>S21.1971</td></tr></tbody></table><p> </p></div></div></div></div><div class="profile-bottom"></div></div></div></div></div> <script type="text/javascript"> jQuery(document).ready(function($) { $('.nomen-content').hide(); $('.nomen-tabs li').hover(function() { $('.nomen-content').show(); 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).change(function() { var country = $(this).val(); if(country == 'US'){ $('.webform-component--city').show(); }else{ $('.edit-submitted-city').val(''); $('.webform-component--city').hide(); } }); $('.tablefield-wrapper table.tablefield.sticky-enabled.tableheader-processed.sticky-table').addClass('table'); }); </script> <style type='text/css'> .embeddedServiceHelpButton .helpButton .uiButton { background-color: #005f9e; font-family: "Arial", sans-serif; max-width: none; } .embeddedServiceHelpButton .embeddedServiceIcon::before { font-size: 2.25em; } .embeddedServiceHelpButton .helpButton .uiButton:hover { background-color: #0097ed; } .embeddedServiceHelpButton .helpButton .uiButton:focus { outline: 1px solid #005290; text-decoration: none; } .embeddedServiceHelpButton .helpButton .uiButton:active { text-decoration: none; } .embeddedServiceHelpButton .helpButton .helpButtonEnabled:hover::before, .embeddedServiceHelpButton .helpButton .helpButtonEnabled:focus::before { background-color: 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background-color: #FFFFFF; padding: 10px; max-height: 50px; border-bottom-left-radius: 8px; border-bottom-right-radius: 8px; } .embeddedServiceInvitationFooter > .embeddedServiceActionButton { font-size: 14px; max-height: 40px; border: none; border-radius: 4px; padding: 10px; margin: 4px; text-align: center; text-decoration: none; display: inline-block; cursor: pointer; } .embeddedServiceInvitationFooter > #acceptInvite { background-color: #005290; color: #FFFFFF; } .embeddedServiceInvitationFooter > #rejectInvite { background-color: #FFFFFF; color: #005290; } </style> <!-- doFind, doCreate and isExactMatch example for a Contact: Find a contact whose Email exactly matches the value provided by the customer in the form If there's no match, then create a Contact record and set it's First Name, Last Name, Email, and Phone to the values provided by the customer --> <!-- <input type="hidden" name="liveagent.prechat.findorcreate.map.doFind:Contact" value="Email,true" /> <input type="hidden" 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embedded_svc.liveAgentAPI.startChat({ directToButtonRouting: { buttonId: "5733t000000cVk9" }, extraPrechatInfo: [], extraPrechatFormDetails: [] }); }; // use this API call to start chat from invitations document.addEventListener('keyup', function(event) { if (event.keyCode == 27) { embedded_svc.inviteAPI.inviteButton.rejectInvite(); }}) })(); </script> <!-- End of Invitations --> <script type='text/javascript' src='https://service.force.com/embeddedservice/5.0/esw.min.js'></script> <script type='text/javascript'> var initESW = function(gslbBaseURL) { embedded_svc.settings.displayHelpButton = true; //Or false embedded_svc.settings.language = ''; //For example, enter 'en' or 'en-US' embedded_svc.settings.defaultMinimizedText = 'Chat now'; //(Defaults to Chat with an Expert) embedded_svc.settings.disabledMinimizedText = 'Leave a message'; //(Defaults to Agent Offline) embedded_svc.settings.autoOpenPostChat = true; //embedded_svc.settings.loadingText = ''; //(Defaults to Loading) //embedded_svc.settings.storageDomain = 'yourdomain.com'; //(Sets the domain for your deployment so that visitors can navigate subdomains during a chat session) // Settings for Chat embedded_svc.settings.directToButtonRouting = function (prechatFormData) { if (prechatFormData[4].value === "Technical Service") return "5733t000000XZAM"; if (prechatFormData[4].value === "Customer Care") return "5733t000000XZAH"; }; embedded_svc.settings.prepopulatedPrechatFields = { OmniURL__c: window.location.href, ChooseaDepartment__c: 'Technical Service' }; //Sets the auto-population of pre-chat form fields embedded_svc.settings.extraPrechatFormDetails = [{ "label":"Source Domain", "value":window.location.href, "transcriptFields":[ "Source_Domain__c" ], "displayToAgent":true }]; //embedded_svc.settings.fallbackRouting = []; //An array of button IDs, user IDs, or userId_buttonId embedded_svc.settings.offlineSupportMinimizedText = 'Leave a message'; //(Defaults to Contact Us) 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type="text/javascript">window.NREUM||(NREUM={});NREUM.info={"beacon":"bam.nr-data.net","licenseKey":"NRJS-dd6144c4de227d09eb4","applicationID":"1022125825","transactionName":"YFxbZkpUXBYEVkYNWVkWbEBRGlsLAVBKSkZfSQ==","queueTime":0,"applicationTime":557,"atts":"TBtYEAJOTxg=","errorBeacon":"bam.nr-data.net","agent":""}</script></body> </html>

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