[{"data":1,"prerenderedAt":-1},["ShallowReactive",2],{"doc-seo-450327-105":3,"detail-sidebar-cat-0-en-105":81,"doc-detail-450327-en":130},{"code":4,"msg":5,"data":6},0,"ok",{"site_id":7,"language":8,"slug":9,"title":10,"keywords":11,"description":12,"schema_data":13,"social_meta":74,"head_meta":76,"extra_data":78,"updated_unix":80},105,"en","visualising-viral-interactions-and-mechanisms-at-the-nanoscale-with-expansion-microscopy","Visualising viral interactions and mechanisms at the nanoscale with expansion microscopy","","Expansion microscopy enables nanoscale imaging of biological specimens with standard optical microscopes by embedding samples in swellable hydrogels. It provides sub-diffraction, typically ~70 nm or better resolution, supports 3D and multi-colour visualisation, and preserves native tissue architecture via isotropic sample enlargement. The workflow covers fixation, biomolecular labelling, hydrogel anchoring, embedding, enzymatic digestion/denaturation, and expansion, with emphasis on compatible fluorophores, efficient linkers, and fluorescence-preserving strategies. The article discusses its potential for virology by enabling cellular and sub-cellular mechanism studies in viruses.",{"@graph":14,"@context":73},[15,34,56],{"@type":16,"itemListElement":17},"BreadcrumbList",[18,23,27,31],{"item":19,"name":20,"@type":21,"position":22},"https://docshare.wps.com","Home","ListItem",1,{"item":24,"name":25,"@type":21,"position":26},"https://docshare.wps.com/document/","Document",2,{"item":28,"name":29,"@type":21,"position":30},"https://docshare.wps.com/document/research-report/","Research & Report",3,{"item":32,"name":10,"@type":21,"position":33},"https://docshare.wps.com/document/visualising-viral-interactions-and-mechanisms-at-the-nanoscale-with-expansion-microscopy/450327/",4,{"url":32,"name":10,"@type":35,"image":36,"author":41,"headline":10,"publisher":44,"fileFormat":47,"inLanguage":8,"description":12,"dateModified":48,"datePublished":49,"encodingFormat":47,"isAccessibleForFree":50,"interactionStatistic":51},"DigitalDocument",{"url":37,"@type":38,"width":39,"height":40},"https://docshare.wps.com/thumbnails/visualising-viral-interactions-and-mechanisms-at-the-nanoscale-with-expansion-microscopy/450327.png","ImageObject",300,407,{"name":42,"@type":43},"Valentina","Person",{"url":19,"name":45,"@type":46},"DocShare","Organization","application/pdf","2026-10-07","2026-09-30",true,{"@type":52,"interactionType":53,"userInteractionCount":55},"InteractionCounter",{"@type":54},"ViewAction",7,{"@type":57,"mainEntity":58},"FAQPage",[59,65,69],{"name":60,"@type":61,"acceptedAnswer":62},"What is expansion microscopy and how does it achieve nanoscale imaging?","Question",{"text":63,"@type":64},"It embeds biological specimens in swellable hydrogels, which isotropically enlarge the sample to reach sub-diffraction resolution, typically on the order of ~70 nm or less. This enables much higher-resolution visualisation using standard optical microscopes.","Answer",{"name":66,"@type":61,"acceptedAnswer":67},"What are the main steps in the standard expansion microscopy workflow?",{"text":68,"@type":64},"The protocol includes fixation, labelling of biomolecules, anchoring chemical linkers, embedding in hydrogel, homogenisation via enzymatic digestion or denaturation, and subsequent expansion by immersion in water or expansion buffer. Each step must be optimised to the sample and target.",{"name":70,"@type":61,"acceptedAnswer":71},"How do fluorophore choice and digestion time affect image quality?",{"text":72,"@type":64},"Fluorophores differ in photo- and chemical stability, and some dye degradation can occur during hydrogel polymerisation. Digestion time must be adjusted for sample type, since excessive digestion can degrade auto-fluorescent proteins and SNAP-tags, while strategies like label retention and signal amplification can preserve fluorescence.","https://schema.org",{"og:url":32,"og:type":75,"og:title":10,"og:site_name":45,"og:description":12},"article",{"robots":77,"canonical":32},"index,follow",{"doc_id":79,"site_id":7},450327,1790771921,{"code":4,"msg":82,"data":83},"success",[84,88,92,96,101,106,110,114,119,122,126],{"id":22,"doc_module":4,"doc_module_name":25,"category_name":85,"show_sort_weight":86,"slug":87},"Story & Novel",90,"story-novel",{"id":26,"doc_module":4,"doc_module_name":25,"category_name":89,"show_sort_weight":90,"slug":91},"Literature",80,"literature",{"id":33,"doc_module":4,"doc_module_name":25,"category_name":93,"show_sort_weight":94,"slug":95},"Exam",70,"exam",{"id":97,"doc_module":4,"doc_module_name":25,"category_name":98,"show_sort_weight":99,"slug":100},5,"Comic",60,"comic",{"id":102,"doc_module":4,"doc_module_name":25,"category_name":103,"show_sort_weight":104,"slug":105},6,"Technology",50,"technology",{"id":55,"doc_module":4,"doc_module_name":25,"category_name":107,"show_sort_weight":108,"slug":109},"Healthcare",40,"healthcare",{"id":111,"doc_module":4,"doc_module_name":25,"category_name":29,"show_sort_weight":112,"slug":113},8,30,"research-report",{"id":115,"doc_module":4,"doc_module_name":25,"category_name":116,"show_sort_weight":117,"slug":118},9,"Religion & Spirituality",20,"religion-spirituality",{"id":117,"doc_module":4,"doc_module_name":25,"category_name":120,"show_sort_weight":117,"slug":121},"World Cup","world-cup",{"id":123,"doc_module":4,"doc_module_name":25,"category_name":124,"show_sort_weight":123,"slug":125},10,"Lifestyle","lifestyle",{"id":127,"doc_module":4,"doc_module_name":25,"category_name":128,"show_sort_weight":97,"slug":129},19,"General","general",{"code":4,"msg":82,"data":131},{"doc_id":79,"user_id":132,"nickname":42,"user_avatar":133,"doc_module":4,"category_id":111,"category_name":29,"doc_title":10,"doc_description":12,"doc_content":134,"file_id":135,"file_url":136,"file_type":137,"file_size":138,"view_count":55,"is_deleted":4,"is_public":22,"is_downloadable":22,"audit_status":22,"page_count":139,"language":140,"language_code":8,"site_id":7,"html_lang":8,"table_of_contents":141,"faqs":142,"seo_title":143,"seo_description":12,"update_tm":144,"read_time":145},13056703020460,"https://ap-avatar.wpscdn.com/avatar/be000253dac470eee5d?_k=1778207105932848923","npj | viruses Review  \n\n| \u003Cbr>[https://doi.org/10.1038/s44298-025-00169-y](https://doi.org/10.1038/s44298-025-00169-y) |  |\n| --- | --- |\n| Visualising viral interactions and mechanisms atthe nanoscale with expansion microscopy\u003Cbr> Check for updates |  |\n| Lisa Övermöhle1,3, Maximilian Baum1,3, Rohan Bhatia2, Karin Byskata1 & Katharina M. Scherer1  |  |\n| Expansion microscopy is a groundbreaking technique that enables nanoscale imaging of biological specimens using standard optical microscopes. By embedding specimens in swellable hydrogels, it achieves sub-diffraction resolution. Compatible with various tissue types, it offers 3D, multi-colour visualisation of cellular and sub-cellular structures. While challenges remain, like sample isotropy and preservation of molecular integrity, expansion microscopy is a transformative tool for cell and neurobiology. Here, we discuss its potential for virology. |  |\n| Principles of expansion microscopy The standard expansion microscopyprotocol involves several key steps(Fig. 1): (1) fixation of the specimen,(2) labelling of biomolecules of interest,(3) anchoring chemical linkers to biomolecules,(4) embedding in a hydrogel,(5) homogenisation through enzymatic digestion or denaturation of the tissue, and(6)subsequent expansion by immersion inwater or an expansion buffer. These steps result in the isotropic enlargement of the sample, preserving the native tissue architecture while enabling visualisation at a significantly higher resolution-typically on the order of 70 nm and less. The exact resolution attained depends on the specific expansion protocol, as different methods yield varying expansion factors (commonly 4x, but protocols achieving 10x and more have been developed) and may also involve iterative rounds of expansion.\u003Cbr>Several practical considerations must be made regarding each individual step. Based on the specific structure of interest, the most suitable reagent should be chosen for fixation. These are usually the same as commonly used for immunocytochemistry, such as methanol and aldehydes (e.g., paraformaldehyde, glutaraldehyde, glyoxal). Cryo-fixation provides the most accurate preservation of biological samples in their native state1; however, it requires more time and skilled handling than conventional chemical fixation methods.\u003Cbr>The selection of fluorescent labels is crucial: some free-radical-induced dye degradation is inevitable during the polymerisation step of forming the hydrogel backbone, but some dyes are more photo-and chemically stable than others. For example, rhodamine-based dyes (e.g., Atto647N) show muchbetter stability than cyanine dyes(e.g., Alexa Fluor647)2 which should best be avoided for expansion microscopy. Autofluorescent and selflabelling proteins can stay intact if digestion is not too prolonged, so the digestion time mustbe adjusted for each sample type like single cells or tissue sections3–6. Although enzymatic homogenisation generally improves | expansion, excessive digestion can degrade these proteins, especially autofluorescent proteins and SNAP-tags. There are further strategies topreserve and enhance fluorescence signals in expanded samples. Label retention might be improved by using small, inert trifunctional probes that bind the target, anchor to the hydrogeland carry afluorophore5. Signal amplification can be achieved using fluorescent nanobodies or tyramide signal amplification7 to boost fluorescence intensity. Post-expansion labelling can improve labelling efficiency due to better epitope accessibility8, often using treatment with sodium dodecyl sulfate(SDS)for long durationsand/or high temperature treatment9, 10 instead of enzymatic digestion.\u003Cbr>The anchoring step requires thorough incubation of the sample with the linker solution to ensure uniform distribution within the sample. Slow polymerisation at lower temperatures during embedding promotes uniform reactions and homogeneity. For expansion microscopy, labelling and anchoring strategies must be co","cbCaiqo3gG8kPHtq","https://ap.wps.com/l/cbCaiqo3gG8kPHtq","pdf",1872178,11,"English","# Principles of expansion microscopy\n## Workflow steps\n## Resolution and expansion factors\n## Practical considerations for sample prep\n## Fluorescent labels and signal preservation\n## Anchoring strategies and biomolecule targeting\n## Imaging biomolecules (proteins, DNA/RNA)","[{\"question\":\"What is expansion microscopy and how does it achieve nanoscale imaging?\",\"answer\":\"It embeds biological specimens in swellable hydrogels, which isotropically enlarge the sample to reach sub-diffraction resolution, typically on the order of ~70 nm or less. This enables much higher-resolution visualisation using standard optical microscopes.\"},{\"question\":\"What are the main steps in the standard expansion microscopy workflow?\",\"answer\":\"The protocol includes fixation, labelling of biomolecules, anchoring chemical linkers, embedding in hydrogel, homogenisation via enzymatic digestion or denaturation, and subsequent expansion by immersion in water or expansion buffer. Each step must be optimised to the sample and target.\"},{\"question\":\"How do fluorophore choice and digestion time affect image quality?\",\"answer\":\"Fluorophores differ in photo- and chemical stability, and some dye degradation can occur during hydrogel polymerisation. Digestion time must be adjusted for sample type, since excessive digestion can degrade auto-fluorescent proteins and SNAP-tags, while strategies like label retention and signal amplification can preserve fluorescence.\"}]","Visualising viral interactions and mechanisms at the nanoscale with expansion microscopy | PDF",1790732905,28]