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The workflow is primarily designed for FFPE tissue sections but can be adapted to other tissue preparations. It closely matches a Standard BioTools protocol, while adding extra guidance and tips for improved execution. The protocol is published as an open-access draft without a DOI, and is intended for careful, step-by-step implementation to maintain signal quality.",{"@graph":69,"@context":122},[70,84,105],{"@type":71,"itemListElement":72},"BreadcrumbList",[73,77,79,82],{"item":74,"name":75,"@type":76,"position":8},"https://docshare.wps.com","Home","ListItem",{"item":78,"name":9,"@type":76,"position":14},"https://docshare.wps.com/document/",{"item":80,"name":35,"@type":76,"position":81},"https://docshare.wps.com/document/healthcare/",3,{"item":83,"name":65,"@type":76,"position":19},"https://docshare.wps.com/document/hyperion-imc-staining-protocol-draft-working/509048/",{"url":83,"name":65,"@type":85,"image":86,"author":91,"headline":65,"publisher":94,"fileFormat":97,"inLanguage":63,"description":67,"dateModified":98,"datePublished":99,"encodingFormat":97,"isAccessibleForFree":100,"interactionStatistic":101},"DigitalDocument",{"url":87,"@type":88,"width":89,"height":90},"https://docshare.wps.com/thumbnails/hyperion-imc-staining-protocol-draft-working/509048.png","ImageObject",300,407,{"name":92,"@type":93},"supergirl","Person",{"url":74,"name":95,"@type":96},"DocShare","Organization","application/pdf","2026-11-11","2026-10-05",true,{"@type":102,"interactionType":103,"userInteractionCount":29},"InteractionCounter",{"@type":104},"ViewAction",{"@type":106,"mainEntity":107},"FAQPage",[108,114,118],{"name":109,"@type":110,"acceptedAnswer":111},"What is the main purpose of this Hyperion IMC staining protocol?","Question",{"text":112,"@type":113},"It provides a staining workflow to detect protein targets using Hyperion imaging mass cytometry. The protocol focuses on preserving and detecting antigen signal throughout the steps.","Answer",{"name":115,"@type":110,"acceptedAnswer":116},"Which sample type is primarily supported, and can it be adapted?",{"text":117,"@type":113},"It is primarily written for working with FFPE tissue sections. The protocol can be adapted to other tissue preparations if needed.",{"name":119,"@type":110,"acceptedAnswer":120},"Why does the protocol emphasize being exact and following steps carefully?",{"text":121,"@type":113},"It states that the protocol is unforgiving if implementation is sloppy. It also notes the approximate cost per stained slide can be high, so careful execution matters.","https://schema.org",{"og:url":83,"og:type":124,"og:title":65,"og:site_name":95,"og:description":67},"article",{"robots":126,"canonical":83},"index,follow",{"doc_id":128,"site_id":62},509048,1791417643,{"code":4,"msg":5,"data":131},{"doc_id":128,"user_id":132,"nickname":92,"user_avatar":133,"doc_module":4,"category_id":34,"category_name":35,"doc_title":65,"doc_description":67,"doc_content":134,"file_id":135,"file_url":136,"file_type":137,"file_size":138,"view_count":29,"is_deleted":4,"is_public":8,"is_downloadable":8,"audit_status":8,"page_count":139,"language":140,"language_code":63,"site_id":62,"html_lang":63,"table_of_contents":141,"faqs":142,"seo_title":143,"seo_description":67,"update_tm":144,"read_time":145,"preview_page_count":4},962088121634,"https://ap-avatar.wpscdn.com/davatar_9964176cb1d06d4a9deccf72a44ae3dc","Sep 18, 2025  \n Hyperion IMC Staining  \nThis protocol is a draft, published without a DOI.  \nMichael Haley1  \n1 Bioimaging Core Facility, Faculty of Biology, Medicine and Health at University of Manchester  \n Michael Haley  \nCreate & collaborate more with a free account  \nEdit and publish protocols, collaborate in communities, share insights through comments, and track progress with run records.  \nCreate free account  \nProtocol Citation: Michael Haley 2025. Hyperion [IMC Staining. protocols.io](IMC Staining. protocols.io) [https://dx.doi.org/](https://dx.doi.org/)  \nLicense: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited  \nProtocol status: Working  \nWe use this protocol and it 's working Created: September 16, 2025  \nLast Modified: September 18, 2025  \nProtocol Integer ID: 227402  \nKeywords: imaging mass cytometry, hyperion, imc, staining protocol, detection of protein target, mass cytometry, using hyperion, standard biotool, ffpe tissue section, protein target, other tissue preparation  \nAbstract  \nThe staining protocol is for detection of protein targets using Hyperion imaging mass cytometry. It is primarily written for working with FFPE tissue sections, though can be adapted to other tissue preparations. It is nearly identical to the protocol provided by Standard BioTools (see references), just with extra guidance and tips.  \nAttachments  \nFLDM􀃂01336􀂫02􀂬 Imagi...  \n658KB  \nHyperion Panel Templ...  \n15KB  \nGuidelines  \nLook for bullet points at the end of steps for specific tips or guidance on those steps.  \nMaterials  \n Xylene. Used pure straight from bottle. Note that two different changes of xylene are used, this is intentional. The first will accumulate more paraffin, with the second therefore being 'cleaner '.  \n Ethanols. Either used pure, or diluted with water to appropriate percentage.  \n Ultra pure or milliQ water. Either should be fine, I would just avoid using distilled water from a normal lab tap unless you are no completely confident in its quality. Historically, I have had doubts about the distilled water in the AV Hill building, especially when they have cleaned the pipes then not washed the excess cleaning reagents out properly!  \n Tris-EDTA solution. 10 mM Tris base, 1 mM EDTA solution, pH 8.5. This is a fairly standard solution used for antigen retrieval, with several manufactures providing their own premade solutions or stocks that should be fine.  \n PAP pen. Any brand should be fine, but once opened they can 'go off ' with time, or dry out.  \n Staining chamber. This should be humidified, usually with a water reservoir of some kind. The entire point is to keep the slides in a moist environment which will prevent even small volumes evaporating from the surface of the slide during the room temperature and over night incubations.  \n 10% BSA stock. We used a 10% BSA stock supplied by ThermoFisher which is also stabilised with something antimicrobial. You can also make up your own stock, but beware that BSA stocks can become contaminated with bacteria, or form aggregates  \n Block buffer. 3% bovine serum albumin 􀂩BSA) in PBS. Dilute from 10% BSA stock.  \n PBS. I would suggest using bottled 1X PBS rather than making your own.  \n 0.1% Triton X100􀃂PBS. Add 500 μl of Triton X􀃂100 to a 500 ml bottle of PBS and shake to mix.  \n Iridium. Stock is Cell-ID Intercalator-Ir—500 µM from Standard Biotools. Used 1􀃥400 in PBS.  \nSafety warnings  \n Anyone familiar with immunohistochemistry should be familiar with most steps in this protocol. However, this protocol rewards being exact and careful, and can be very unforgiving if you are sloppy, or do not follow the protocol. The approximate cost to stain one slide with an entire panel is £300􀃂400! Therefore, in even small pilot experiments can add up fairly quickly.  \nBefore start  \n Make sure ","cbCaiqEpMNZs2UVx","https://ap.wps.com/l/cbCaiqEpMNZs2UVx","pdf",899111,11,"English","# Materials\n## Safety warnings\n# Before start\n## Preparation 3h\n## Note","[{\"question\":\"What is the main purpose of this Hyperion IMC staining protocol?\",\"answer\":\"It provides a staining workflow to detect protein targets using Hyperion imaging mass cytometry. The protocol focuses on preserving and detecting antigen signal throughout the steps.\"},{\"question\":\"Which sample type is primarily supported, and can it be adapted?\",\"answer\":\"It is primarily written for working with FFPE tissue sections. The protocol can be adapted to other tissue preparations if needed.\"},{\"question\":\"Why does the protocol emphasize being exact and following steps carefully?\",\"answer\":\"It states that the protocol is unforgiving if implementation is sloppy. It also notes the approximate cost per stained slide can be high, so careful execution matters.\"}]","Hyperion IMC Staining - Protocol Draft - Working | PDF",1791233976,28]