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Innovative Uses of RNAscope | RNAScope

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id="block-system-main" class="block block-system clearfix"><div class="content"><div id="node-70912" class="node node-innovation node-promoted" about="/innovations-rnascope" typeof="sioc:Item foaf:Document"><div class="tab-content"><div class="order-container container home-tab tab-pane fade in active" id="innovation-container"><div class="spotlights-container"><div class="spotlight-top"><div class="spotlight-top-inner"><div class="spotlight-title">Innovative Uses of RNAscope</div><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>Read about reseachers who take innovative approaches with RNAscope™ protocols to advance their research. If you would like to try one of these customer-demonstrated applications, please note that they are non-standard applications and therefore, a careful read through of the publications/materials is highly recommended. Our technical support team is here and will make best efforts to support you.</p></div></div></div></div></div><div class="spotlight-bottom"><div class="spotlight-middle" style="background-image:url(https://acdbio.com/sites/default/files/ACD_HIV-Innovation-BKG.jpg)"><div class="spotlight-middle-inner"><div class="innovation-sub-title">Customer demonstrated application:</div><div class="spotlight-middle-title">SIV/HIV RNA & DNA detection</div> <a href="#" class="see-more wish-more-link2">Read More</a></div></div></div><div class="wish-more-content2"><div class="wish-more-desp desp1"><div class="wish-more-inner"><p style="font-size: 18px;margin-bottom: 20px;">ACD congratulates the team of scientists, led by Jacob D. Estes on their publication <a href="https://paijournal.com/index.php/paijournal/article/view/100/43"><strong>Defining HIV and SIV Reservoirs in Lymphoid Tissues</strong></a> DOI 10.20411/pai.v1i1.100.</p><p>The authors aimed to track and discriminate HIV-1/SIV viral reservoirs within tissue compartments and thus applied a 'specific and sensitive next-generation <em>in situ</em> hybridization approach.'&nbsp;The authors demonstrated:</p><p>1) that an optimized next-generation ISH platform RNAscope Assay for the rapid detection of vRNA (with results obtained within 1 day) has sufficient sensitivity to reliably detect single virions in B cell follicles (BCF) in FFPE tissue sections,</p><p>2) that an approach for the detection of vDNA <em>in situ</em> (referred to as DNAscope) reliably and readily detects vDNA+ cells, and</p><p>3) that they have developed an <em>in situ</em> method to simultaneously visualize vRNA and vDNA in the same tissue section and thereby identify transcriptionally latent infections (vDNA+/vRNA- cells) in lymphoid tissues.</p><p>The team of researchers then applied “these new, highly sensitive <em>in situ</em> hybridization approaches" to lymphoid tissues samples from macaques prior to and during combination anti-retroviral therapy (cART) and documented the importance of BCFs in active, latent&nbsp;and persistent infections during treatment. Authors concluded that “these data underscore the utility of new and sensitive ISH tools that can provide additional insight into viral persistence, reservoir establishment, tissue compartmentalization&nbsp;and reservoir phenotype in the local<em> in vivo </em>tissue environment.”</p><p><a class="watch-link" href="https://paijournal.com/index.php/paijournal/article/view/100/43" style="line-height: 20.8px;" target="_blank">Read the Publication</a></p><p><span style="font-size:18px;"><strong>Contact ACD to get&nbsp;started</strong></span></p><p>Visualization of viral DNA as described above is a non-standard, modified RNAscope<sup><sup>®</sup>&nbsp;</sup>protocol. &nbsp;Please familiarize yourself with the methodology described. The ACD technical&nbsp;support team will provide the best support for you. &nbsp;Please note, as yet, there are&nbsp;no experimental data demonstrating the extension of this protocol to other&nbsp;species, other probes or other tissue types not described in the publication. &nbsp;</p><p><a class="watch-link" href="https://acdbio.com/contact" style="line-height: 20.8px;" target="_blank">Contact Us</a></p></div></div><div class="wish-more-desp desp2"><div class="wish-more-inner"><p><span style="font-size:18px;"><strong>What to order from ACD </strong></span></p><p>Your account executive will ensure completeness of your order. &nbsp;Standard accessories needed for any of the three protocols below include:</p><p style="line-height: 20.8px;">1. ACD HybEZ Hydridization System with ACD EZBatch&nbsp;Cat. No. <a href="/search/site/321461/cms/product">321461</a>&nbsp;110V&nbsp;or&nbsp;<span style="line-height: 20.8px;">Cat. No.&nbsp;<a href="/search/site/321462/cms/product">321462</a>&nbsp;220V</span></p><p style="line-height: 20.8px;"><span style="line-height: 20.8px;">2. ImmEdge Hydrophobic Barrier Pen Cat. No. <a href="/search/site/310018/cms/product">310018</a></span></p><p style="line-height: 20.8px;"><span style="line-height: 20.8px;">3. Control probes: &nbsp;Negative control probe and species-specific positive control probes are also required&nbsp;</span></p><p style="line-height: 20.8px;">&nbsp;</p><p><strong>To detect </strong><strong style="line-height: 20.8px;">and visualize</strong><strong>&nbsp;HIV-1 AND SIV viral RNA </strong><i><strong>in situ,</strong> the authors used <u>standard</u> RNAscope Assay Protocol and anti-sense probes outlined in Supplemental Table 2</i>. Additionally the following materials from ACD&nbsp;are required:</p><p>1. &nbsp;RNAscope 2.5 HD Red Kit: Cat. No.&nbsp;<a href="/search/site/322350/cms/product">322350</a>&nbsp;</p><p>2. EcoMount Mounting Medium Cat. No. <a href="/search/site/320409/cms/product">320409</a></p><p>&nbsp;</p><p><strong>To detect and visualize&nbsp;HIV-1 AND SIV viral DNA <em>in situ</em>,</strong>&nbsp;<i style="line-height: 20.8px;">the authors used <u>modified</u>&nbsp;RNAscope Assay Protocol and sense probes outlined in Supplemental Table 2</i><span style="line-height: 20.8px;">. Additionally the following materials from ACD&nbsp;are required:</span></p><p>1.RNAscope 2.5 HD Brown Kit Cat. No.&nbsp;<a href="/search/site/322300/cms/product">322300</a></p><p style="line-height: 20.8px;">&nbsp;</p><p><strong style="line-height: 20.8px;">To detect and visualize both viral RNA and viral DNA <em>in situ</em>, in a single assay&nbsp;</strong><i style="line-height: 20.8px;">the authors used <u>modified</u>&nbsp;RNAscope Assay Protocol both sense and anti-sense probes outlined in Supplemental Table 3</i><span style="line-height: 20.8px;">. The following materials&nbsp;from ACD are also required:</span></p><p>1. RNAscope 2.5 HD Duplex Kit Cat. No.&nbsp;<a href="/search/site/322430/cms/product">322430</a></p><p>2. VectaMount Permanent Mounting Medium Cat. No.<a href="/search/site/321584/cms/product">321584</a></p><p style="line-height: 20.8px;"><a>&nbsp;</a></p></div></div></div><div class="spotlight-bottom"><div class="spotlight-middle" style="background-image:url(https://acdbio.com/sites/default/files/innovations_0.jpg)"><div class="spotlight-middle-inner"><div class="innovation-sub-title">Customer demonstrated application:</div><div class="spotlight-middle-title">WISH Whole-Mount in situ Hybridization</div> <a href="#" class="see-more wish-more-link">Read More</a></div></div></div><div class="wish-more-content"><div class="wish-more-desp desp1"><div class="wish-more-inner"><p>Our ACD team are&nbsp;excited that Theresa Gross-Thebing, Azadeh Paksa and Erez Raz at&nbsp;Münster&nbsp;University, Germany have driven the RNAscope™ technology into a new arena - establishing, optimizing&nbsp;and applying RNAscope™ Assay to whole mount embryonic zebrafish samples.&nbsp;Although RNAscope™ assay may&nbsp;be applied to any gene and any species, appropriate sample preparation is essential.</p><p>Contact the ACD support team at <a href="mailto:support.ACD@bio-techne.com">support.ACD@bio-techne.com</a> for detailed support for&nbsp;the of RNAscope™ Assay.</p><p>Theresa Gross-Thebing and Azadeh Paksa have generously volunteered to answer questions specific to the preparation of whole-mount embryonic samples. &nbsp;Email them using&nbsp;<u><span style="color:#008080">RNAscope4zebrafish@bio-techne.com</span></u> but please note, the response time may take up to 1 week.</p><p>A Tips &amp; Tricks and FAQ section surrounding WISH is under development, please check back with us for updates.</p><p><a class="watch-link" href="http://www.acdbio.com/science/scientific-resources/webinars" target="_blank">Watch the Webinar</a></p></div></div><div class="wish-more-desp desp2"><div class="wish-more-inner"><h4>&nbsp; WHAT TO ORDER TO GET STARTED?</h4><ol><li><strong><strong>RNAscope™ p</strong>ositive control probe: </strong>&nbsp;use zebrafish-specific housekeeping gene for positive control. The Erez Raz Lab used zebrafish myoD gene in channel 2,&nbsp; <u><a href="http://www.acdbio.com/search/site/search&amp;keywords=402461-C2/">CAT # 402461-C2&nbsp; Dr-myod1-C2.</a></u>&nbsp; Alternatively, we also offer CAT# 427301-C2 Dr-gsc-C2 and CAT# 428601 Dr-odc1 (channel 1)</li><li><strong>RNAscope™ negative control probe:</strong> <u><a href="http://www.acdbio.com/search/site/search&amp;keywords=310043+/">CAT # 310043 Negative Control Probe-DapB</a></u></li><li><strong>RNAscope™ Target Probes: &nbsp;</strong>&nbsp;ACD currently offers 33 zebrafish-specific probes (www.acdbio.com/probesearch).&nbsp; New or custom target probes can be designed and manufactured within 2 weeks.&nbsp;</li><li><strong>RNAscope™ Assay Reagent Kit:</strong>&nbsp; The Erez Raz Lab used<strong> </strong><a href="http://www.acdbio.com/products/rnascope-assays/rnascope-fl">CAT # 320850 RNAscope™ Multiplex Fluorescent Reagent Kit</a>, comprise of&nbsp;&nbsp;three separate components that are also available separately:&nbsp;</li></ol><ul><li>CAT # 310091 Wash Buffers</li><li>CAT # 320842 RNAscope™ Pretreatment Kit&nbsp;</li><li>CAT # 320851 RNAscope™ Multiplex Fluorescent Detection Kit</li></ul><p>A multispectral microscopy imaging system is required for visualization. You will be able to investigate&nbsp;up to 4 genes simultaneously with this assay.&nbsp;&nbsp; Additionally, ACD offers three chromogenic assays; however those assays have not been applied to whole-mount zebrafish embryonic samples.</p></div></div></div><div class="spotlight-bottom"><div class="spotlight-middle" style="background-image:url(https://acdbio.com/sites/default/files/researcher.jpg)"><div class="spotlight-middle-inner"><div class="innovation-sub-title">Customer demonstrated application:</div><div class="spotlight-middle-title">Researchers Successfully Apply RNAscope ISH to 25-year-old FFPE Samples</div> <a href="#" class="see-more wish-more-link1">Read More</a></div></div></div><div class="wish-more-content1"><div class="wish-more-desp desp1"><div class="wish-more-inner"><h3>Innovations in the Spotlight:</h3><h3>Erasmus MC Researchers Successfully Applied RNAscope<sup><strong style="line-height: 20.8px;"><sup>®</sup></strong></sup> ISH to 25 year old FFPE Samples</h3><p>&nbsp;</p><p>You may&nbsp;be surprised to know that your archived FFPE samples may <strong>not</strong> be too old for RNAscope™ <em>in situ</em> hybridization (ISH)!&nbsp;Our R&amp;D team routinely detects RNA transcripts in three year old FFPE tissue specimens and some of our customers have applied RNAscope to much older tissues. &nbsp;Read below, data from one of our customers where RNAscope ISH was successfully used in tissue specimens older than 25 years.</p><p>Success of RNAscope™ with older FFPE samples depends on many factors including sample fixation, quality&nbsp;and storage.&nbsp;ACD cannot guarantee results for samples outside of our protocol recommendations. Please read&nbsp;<a href="http://www.acdbio.com/ffpe#faq" style="box-sizing: border-box; color: black; text-decoration: none;"><strong>FAQs for RNAscope<strong><sup>®</sup></strong>&nbsp;assay on archived FFPE samples&nbsp;</strong></a>section below for more information<strong>.</strong></p><p><strong>ACD Customers at Erasmus MC Push the Limits within RNAscope™ FFFPE Protocol</strong></p><p>Researchers<strong>&nbsp;Marije Hoogland and Esther Verhoef</strong>&nbsp;(<a href="http://www.erasmusmc.nl/pathologie/?lang=en" style="box-sizing: border-box; color: black; text-decoration: none;" target="_blank">Department of Pathology, Erasmus MC Rotterdam, the Netherlands</a>) have conducted a retrospective study and successfully performed RNAscope™ <em>in situ </em>hybridization on samples containing human metastases of prostate cancer in a lymph node. The samples were collected in 1987, 1988 and 1989 and analyzed in 2014 with RNAscope™ probe targeting UBC gene. At 25 to 27 years old, these are the oldest samples we know of that have been used with RNAscope™ <em>in situ</em> hybridization. The team followed ACD protocols and user manuals.</p></div></div><div class="wish-more-desp desp2"><div class="wish-more-inner"><h3>FAQs for RNAscope™ Assay on archived FFPE samples</h3><p><strong>How can I ensure that my FFPE tissue specimens will work with RNAscope™ FFPE Assay?</strong></p><p>We have conducted extensive assay development to make sure that the RNAscope™ FFPE assay will work with a wide range of tissue types.</p><ul><li>If your tissue specimens are processed according to the Sample Preparation Guide in the User Manual and they are prepared within the last 3 years, you should expect successful staining for positive control probe using our assay procedure.</li><li>If your specimens are prepared differently from our recommended procedure, RNAscope™ can still be used but may require assay optimization for some procedural steps. Please refer to our&nbsp;<a href="http://www.acdbio.com/technical-support/getting-started" style="box-sizing: border-box; color: black; text-decoration: none;">getting started page</a>&nbsp;for protocol and sample guidelines and optimization.</li><li>Ensure that the assay is run according to RNAscope™ protocol.</li></ul><p><strong>What are the recommended RNAscope™ sample storage and pretreatment conditions for FFPE samples?</strong></p><p>ACD sample storage requirements can be found in our RNAscope™&nbsp;<a href="http://www.acdbio.com/technical-support/downloads/technical-doc/rnascope-ffpe-tissue-specimen-prep/" style="box-sizing: border-box; color: black; text-decoration: none;">Sample Preparation and Pretreatment Guide for FFPE Tissue, PART 1</a>. The recommended pretreatment conditions for FFPE samples are based on tissue that has been fixed for 16-32 hrs at room temperature in 10% NBF.</p><p><strong>What should I do if I don’t know whether the FFPE tissue specimen is appropriately fixed and processed?</strong></p><p>Determine your samples' RNA quality following optimization recommendations on our&nbsp;<a href="http://www.acdbio.com/technical-support/getting-started" style="box-sizing: border-box; color: black; text-decoration: none;">getting started page&nbsp;</a>(www.acdbio.com/go).</p></div></div><div class="profile innovation-slider"><div class="loader-image-gallery"></div><div id="film_roll" class="sample-slider"><div> <img class="img-slider" src="https://acdbio.com/sites/default/files/FFPE1.jpg"/><div class="slider-below-text"><p>Figure 1: Punctate red dots show expression of UBC gene in Human prostate cancer samples from lymph node. RNAscope Chromogenic Red assay applied to FFPE&nbsp; sample from 1989 , 400x magnification.</p></div></div><div> <img class="img-slider" src="https://acdbio.com/sites/default/files/FFPE2.jpg"/><div class="slider-below-text"><p>Figure 2: Expression of UBC gene (shown as punctate red dots) in human prostate cancer samples from lymph node. RNAscope Chromogenic Red Assay applied to FFPE samples from 1989, 400x magnification.</p></div></div><div> <img class="img-slider" src="https://acdbio.com/sites/default/files/FFPE3.jpg"/><div class="slider-below-text"><p>Figure 3: Expression of UBC gene (shown as punctate red dots) in human prostate cancer samples from lymph node. RNAscope Chromogenic Red Assay applied to FFPE samples from 1987, 400x magnification.</p></div></div><div> <img class="img-slider" src="https://acdbio.com/sites/default/files/FFPE4.jpg"/><div class="slider-below-text"><p>Figure 4: Expression of UBC gene (shown as punctate red dots) in human prostate cancer samples from lymph node. RNAscope Chromogenic Red assay applied to FFPE samples from 1989, 400x magnification.</p></div></div></div></div></div></div></div></div></div> <script type="text/javascript"> jQuery(document).ready(function($) { // jQuery(function() { // fr = new FilmRoll({ // configure_load: true, // container: '#film_roll', // pager:false, // scroll:false, // }); // }); $('.wish-more-link').click(function(event) { event.preventDefault(); if($(this).hasClass('active')){ $(this).removeClass('active'); $(this).html('Read More'); }else{ $(this).addClass('active'); $(this).html('Read Less'); } $('.wish-more-content').slideToggle(); }); $('.wish-more-link1').click(function(event) { event.preventDefault(); if($(this).hasClass('active')){ $(this).removeClass('active'); $(this).html('Read More'); $('.wish-more-content1').slideUp(); }else{ $(this).addClass('active'); $(this).html('Read Less'); $('.wish-more-content1').slideDown('slow/400/fast', function() { jQuery(function() { fr = new FilmRoll({ configure_load: true, container: '#film_roll', pager:false, scroll:false, }); 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action="https://proteinsimple--obtfull.my.salesforce.com/servlet/servlet.WebToCase?encoding=UTF-8" id="web-to-case1" method="POST"><div class="row"><div class="col-md-6"> <label for="fname">First Name</label> <input id="fname" maxlength="80" name="fname" size="20" type="text"></div><div class="col-md-6"> <label for="lname">Last Name</label> <input id="lname" maxlength="80" name="lname" size="20" type="text"></div></div><div class="row"><div class="col-md-6"> <label for="email">Email</label> <input id="email" maxlength="80" name="email" size="20" type="text"></div><div class="col-md-6"> <label for="phone">Phone</label> <input id="phone" maxlength="40" name="phone" size="20" type="text"></div></div><div class="row"><div class="col-md-6"> <label for="company">Institution / Company</label> <input id="company" maxlength="80" name="company" size="20" type="text"></div></div><div class="row"><div class="col-md-12"> <label for="description">Message</label> <textarea name="description"></textarea> </div></div><div class="row captcha"><div class="col-lg-12 col-md-12 col-sm-12 col-xs-12"> <label>Type the characters that you see in the picture below</label> <input type="text" id="PageCaseCaptcha" disabled /> <a class="btnrefresh" onclick="DrawCaptcha();"></a></div></div><div class="row captcha"><div class="col-lg-12 col-md-12 col-sm-12 col-xs-12"> <input type="text" class="case_captcha" id="PageCaseInput" name="case_captcha"/><div id="pagecasecaptcha-error"></div></div></div> <input type="submit" name="submit" style="float:left;" onclick="ValidCaptcha('pagecase');"><div class="clear"></div></form></div></div><div class="needhelp-tab-container tab-pane fade" id="contactinfo"><div class="supportform-con contact-info"><div class="row"><h3>Advanced Cell Diagnostics</h3><p>Our new headquarters office starting May 2016:</p><p>7707 Gateway Blvd.  <br />Newark, CA 94560<br />Toll Free: 1 (877) 576-3636<br />Phone: (510) 576-8800<br />Fax: (510) 576-8798</p></div><div class="row"><p> </p><div class="row"><h3>Bio-Techne</h3><p>19 Barton Lane  <br />Abingdon Science Park<br />Abingdon<br />OX14 3NB<br />United Kingdom<br />Phone 2: +44 1235 529449<br />Fax: +44 1235 533420</p><p> </p></div><div class="row"><h3>Advanced Cell Diagnostics China</h3><p>20F, Tower 3,<br />Raffles City Changning Office,<br />1193 Changning Road, Shanghai 200051</p><p>021-52293200<br /><a href="mailto:info.cn@bio-techne.com">info.cn@bio-techne.com</a><br />Web: <a href="http://www.acdbio.com/cn">www.acdbio.com/cn</a></p></div><div class="row"><p>For general information: <a href="mailto:Info.ACD@bio-techne.com">Info.ACD@bio-techne.com</a><br />For place an order: <a href="mailto:order.ACD@bio-techne.com">order.ACD@bio-techne.com</a><br />For product support: <a href="mailto:support.ACD@bio-techne.com">support.ACD@bio-techne.com</a><br />For career opportunities: <a href="mailto:hr.ACD@bio-techne.com">hr.ACD@bio-techne.com</a></p></div></div><div class="row" style="border-bottom: none;"><div class="col-md-6"> <a href="/about/distributors" class="meet-out-sales-team" title="See Distributors">See Distributors</a></div></div></div></div><div class="needhelp-tab-container tab-pane fade" id="contactpharma"><div class="supportform-con"><div class="contact-tab-form supportform-con"><form id="mktoForm_9817"></form> <script>MktoForms2.loadForm("//info.bio-techne.com", "584-XSF-168", 9817);</script> </div></div></div></div></div></div></div></div></div></div> <script type="text/javascript"> jQuery(document).ready(function($) { $("#web-to-case1").validate({ // Specify the validation rules rules: { fname: "required", lname: "required", phone:"required", email: { required: true, email: true }, company: "required", case_captcha: "required" }, // Specify the validation error messages messages: { fname: "Please enter your First Name", lname: "Please enter your Last Name", email: "Please enter your email address", phone: "Please enter your 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