{"id":3418,"date":"2021-09-27T09:40:30","date_gmt":"2021-09-27T09:40:30","guid":{"rendered":"https:\/\/www.embl.org\/about\/info\/imaging-centre\/?page_id=3418"},"modified":"2022-06-20T07:53:01","modified_gmt":"2022-06-20T07:53:01","slug":"stimulated-emission-depletion-sted-microscopy","status":"publish","type":"page","link":"https:\/\/www.embl.org\/about\/info\/imaging-centre\/stimulated-emission-depletion-sted-microscopy\/","title":{"rendered":"Stimulated Emission Depletion (STED) Microscopy"},"content":{"rendered":"\n<div class=\"vf-grid | vf-grid__col-3\"><div class=\"vf-grid__col--span-2\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<p>If you want to know how STED works in comparison to other super-resolution technologies we offer &#8211; read more <a href=\"https:\/\/www.embl.org\/about\/info\/imaging-centre\/super-resolution-microscopy\/\">here<\/a>.<\/p>\n\n<\/div>\n<\/div>\n\n\n<div class=\"\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<\/div>\n<\/div>\n<\/div>\n\n\n\n<div class=\"vf-grid | vf-grid__col-1\"><div class=\"\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<div class=\"vf-tabs\"><ul class=\"vf-tabs__list\" data-vf-js-tabs=\"true\"><li class=\"vf-tabs__item\"><a class=\"vf-tabs__link\" href=\"#vf-tabs__section-f7a77b3f-ce92-44e0-9cf6-190701f505a3\" data-vf-js-location-nearest-activation-target=\"\">STELLARIS 8 STED Falcon<\/a><\/li><li class=\"vf-tabs__item\"><a class=\"vf-tabs__link\" href=\"#vf-tabs__section-303f0718-4694-4609-ae98-f62de8c4d8e6\" data-vf-js-location-nearest-activation-target=\"\">STELLARIS STED<\/a><\/li><\/ul><div class=\"vf-tabs-content\" data-vf-js-tabs-content=\"true\">\n<section class=\"vf-tabs__section\" id=\"vf-tabs__section-f7a77b3f-ce92-44e0-9cf6-190701f505a3\"><h2>STELLARIS 8 STED Falcon<\/h2>\n<div class=\"vf-grid | vf-grid__col-3\"><div class=\"vf-grid__col--span-2\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<p>STELLARIS 8 STED with&nbsp;TauSTED&nbsp;from Leica Microsystems allows the study of multiple dynamic events simultaneously and investigation of molecular relationships and mechanisms within the cellular context.<\/p>\n\n\n\n<p>FALCON (FAst&nbsp;Lifetime&nbsp;CONtrast) includes phasors for quantitative FLIM analysis. It is a fully integrated solution for fluorescence lifetime imaging (FLIM) and enables video-rate image acquisition for rapid kinetic studies in live cells.&nbsp;<\/p>\n\n\n\n<hr class=\"vf-divider\">\n\n\n\n<p><strong>Features<\/strong><\/p>\n\n\n\n<ul class=\"wp-block-list\"><li>Resolving molecular relationships in specimens<\/li><li>Multiple simultaneous events can be studied at the nanoscale<\/li><li>2D and 3D STED imaging <\/li><li>Significantly reduced light doses due to&nbsp;TauSTED<\/li><li>Advanced STED-FLIM options with STED and FALCON (full FLIM quantitative tools, phasor FLIM, species separation based on phasor analysis)<\/li><li>Advanced STED-FCS options<\/li><li>Fast validation of results on a single platform, from confocal to super-resolution LIGHTNING and STED<\/li><li>With FALCON it is possible to:<ul><li>follow fast molecular interactions via FLIM-FRET (F\u00f6rster&nbsp;resonance energy transfer).<\/li><li>use biosensors to detect microenvironmental changes, such as pH or ion concentration.<\/li><li>apply lifetime contrast to separate multiple fluorophores.<\/li><li>do analysis using phasor FLIM provides a 2D visualisation of lifetime components. With FLIM phasors you can follow microenvironmental changes, select components to multiplex signal (phasor separation), and determine FRET efficiency.<\/li><\/ul><\/li><\/ul>\n\n\n\n<p><strong>Specifications<\/strong><\/p>\n\n\n\n<ul class=\"wp-block-list\"><li>Five spectrally tunable Power&nbsp;HyD&nbsp;sensitive photon counting detectors (2&nbsp;HyD&nbsp;S, 2&nbsp;HyD&nbsp;X, 1&nbsp;HyD&nbsp;R)<\/li><li>Confocal: tunable White Light Lasers (WLL) 440-790 nm, 405 nm STED depletion: 592 nm, 660 nm, 775 nm<\/li><li>8kHz Resonant Scanner<\/li><li>Total system dead-time: 1.5 ns<\/li><li>TauSTED: Tunable resolution based on lifetime (depending on sample and fluorophore: &lt;30 nm (lateral) and &lt;100 nm (axial). Automatic lifetime-based background suppression algorithm. Light dose reduction (WLL excitation) for all STED lines (592, 660, 775 nm). Available for 2D and 3D STED in live and in fixed specimens, also for multicolor applications. Automated workflow integrated in the LAS X software.<\/li><li>The STED WHITE glycerol and water objective lenses with&nbsp;motCORR&nbsp;technology provide adaptive optical correction for aberrations introduced by sample inhomogeneities and refractive-index mismatch. The STED WHITE objective lenses provide a working distance of 300&nbsp;\u00b5m:<ul><li>HC PL APO 86x\/1.20 W&nbsp;motCORR&nbsp;STED WHITE<\/li><li>HC PL APO 93x\/1.30 GLYC&nbsp;motCORR&nbsp;STED WHITE<\/li><li>HC PL APO 100x\/1.40 OIL STED WHITE<\/li><\/ul><\/li><\/ul>\n\n\n\n<div style=\"height:19px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n<\/div>\n<\/div>\n\n\n<div class=\"\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<figure class=\"vf-figure wp-block-image size-large\"><img loading=\"lazy\" decoding=\"async\" width=\"541\" height=\"1024\" class=\"vf-figure__image\" src=\"https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-content\/uploads\/2021\/12\/Stellaris-8-STED-Falcon-figure-new-541x1024.jpg\" alt=\"Credit: Timo Zimmermann\/EMBL.\" class=\"wp-image-4424\" srcset=\"https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-content\/uploads\/2021\/12\/Stellaris-8-STED-Falcon-figure-new-541x1024.jpg 541w, https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-content\/uploads\/2021\/12\/Stellaris-8-STED-Falcon-figure-new-158x300.jpg 158w, https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-content\/uploads\/2021\/12\/Stellaris-8-STED-Falcon-figure-new.jpg 569w\" sizes=\"auto, (max-width: 541px) 100vw, 541px\" \/><figcaption class=\"vf-figure__caption\">U2OS cells: FLIM images under confocal and STED conditions of nuclear pore protein Nup96 and their corresponding phasor plots of lifetime information. TauSTED image of the same cell. Credit: Timo Zimmermann\/EMBL.<\/figcaption><\/figure>\n\n\n\n<figure class=\"vf-figure wp-block-image size-large\"><img loading=\"lazy\" decoding=\"async\" width=\"1024\" height=\"683\" class=\"vf-figure__image\" src=\"https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-content\/uploads\/2021\/12\/IC_Equipment_003-Edit-1024x683.jpg\" alt=\"Credit: Stuart Bailey\/EMBL.\" class=\"wp-image-4422\" srcset=\"https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-content\/uploads\/2021\/12\/IC_Equipment_003-Edit-1024x683.jpg 1024w, https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-content\/uploads\/2021\/12\/IC_Equipment_003-Edit-300x200.jpg 300w, https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-content\/uploads\/2021\/12\/IC_Equipment_003-Edit-768x512.jpg 768w, https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-content\/uploads\/2021\/12\/IC_Equipment_003-Edit.jpg 1200w\" sizes=\"auto, (max-width: 1024px) 100vw, 1024px\" \/><figcaption class=\"vf-figure__caption\">Leica STELLARIS 8 STED Falcon. Credit: Stuart Bailey\/EMBL. <\/figcaption><\/figure>\n\n<\/div>\n<\/div>\n<\/div>\n<\/section>\n\n\n\n<section class=\"vf-tabs__section\" id=\"vf-tabs__section-303f0718-4694-4609-ae98-f62de8c4d8e6\"><h2>STELLARIS STED<\/h2>\n<div class=\"vf-grid | vf-grid__col-3\"><div class=\"vf-grid__col--span-2\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<p>STELLARIS STED equipped with&nbsp;TauSTED&nbsp;from Leica Microsystems allows the study of multiple dynamic events simultaneously and investigation of molecular relationships and mechanisms within the cellular context.<\/p>\n\n\n\n<hr class=\"vf-divider\">\n\n\n\n<p><strong>Features<\/strong><\/p>\n\n\n\n<ul class=\"wp-block-list\"><li>Resolving molecular relationships in specimens<\/li><li>Multiple simultaneous events can be studied at the nanoscale<\/li><li>2D and 3D STED imaging <\/li><li>Significantly reduced light doses due to&nbsp;TauSTED<\/li><li>Fast validation of results on a single platform, from confocal to super-resolution LIGHTNING and STED<\/li><\/ul>\n\n\n\n<p><strong>Specifications<\/strong><\/p>\n\n\n\n<ul class=\"wp-block-list\"><li>Five spectrally tunable Power&nbsp;HyD&nbsp;sensitive photon counting detectors (2&nbsp;HyD&nbsp;S, 2&nbsp;HyD&nbsp;X, 1&nbsp;HyD&nbsp;R)<\/li><li>Confocal: tunable White Light Lasers (WLL) 440-790 nm, 405 nm STED depletion: 592 nm, 660 nm, 775 nm<\/li><li>8kHz Resonant Scanner<\/li><li>Total system dead-time: 1.5 ns<\/li><li>TauSTED: Tunable resolution based on lifetime (depending on sample and fluorophore: &lt;30 nm (lateral) and &lt;100 nm (axial). Automatic lifetime-based background suppression algorithm. Light dose reduction (WLL excitation) for all STED lines (592, 660, 775 nm). Available for 2D and 3D STED in live and in fixed specimens, also for multicolor applications. Automated workflow integrated in the LAS X software.<\/li><li>The STED WHITE glycerol and water objective lenses with&nbsp;motCORR&nbsp;technology provide adaptive optical correction for aberrations introduced by sample inhomogeneities and refractive-index mismatch. The STED WHITE objective lenses provide a working distance of 300&nbsp;\u00b5m: <ul><li>HC PL APO 86x\/1.20 W&nbsp;motCORR&nbsp;STED WHITE<\/li><li>HC PL APO 93x\/1.30 GLYC&nbsp;motCORR&nbsp;STED WHITE<\/li><li>HC PL APO 100x\/1.40 OIL STED WHITE<\/li><\/ul><\/li><\/ul>\n\n\n\n<div style=\"height:47px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n<\/div>\n<\/div>\n\n\n<div class=\"\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<figure class=\"vf-figure wp-block-image size-large\"><img loading=\"lazy\" decoding=\"async\" width=\"512\" height=\"512\" class=\"vf-figure__image\" src=\"https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-content\/uploads\/2021\/12\/TauSTED-overview-insert-Stellaris-5-STED.jpg\" alt=\"Credit: Timo Zimmermann\/EMBL\" class=\"wp-image-4428\" srcset=\"https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-content\/uploads\/2021\/12\/TauSTED-overview-insert-Stellaris-5-STED.jpg 512w, https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-content\/uploads\/2021\/12\/TauSTED-overview-insert-Stellaris-5-STED-300x300.jpg 300w, https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-content\/uploads\/2021\/12\/TauSTED-overview-insert-Stellaris-5-STED-150x150.jpg 150w\" sizes=\"auto, (max-width: 512px) 100vw, 512px\" \/><figcaption class=\"vf-figure__caption\">U2OS cells: Comparison of confocal and TauSTED images of nuclear pore protein Nup96. Credit: Timo Zimmermann\/EMBL.<\/figcaption><\/figure>\n\n\n\n<figure class=\"vf-figure wp-block-image size-large\"><img loading=\"lazy\" decoding=\"async\" width=\"1024\" height=\"683\" class=\"vf-figure__image\" src=\"https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-content\/uploads\/2021\/12\/IC_Equipment_047-Edit-1024x683.jpg\" alt=\"Credit: Stuart Bailey\/EMBL.\" class=\"wp-image-4426\" srcset=\"https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-content\/uploads\/2021\/12\/IC_Equipment_047-Edit-1024x683.jpg 1024w, https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-content\/uploads\/2021\/12\/IC_Equipment_047-Edit-300x200.jpg 300w, https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-content\/uploads\/2021\/12\/IC_Equipment_047-Edit-768x512.jpg 768w, https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-content\/uploads\/2021\/12\/IC_Equipment_047-Edit.jpg 1200w\" sizes=\"auto, (max-width: 1024px) 100vw, 1024px\" \/><figcaption class=\"vf-figure__caption\">Leica STELLARIS STED. Credit: Stuart Bailey\/EMBL.<\/figcaption><\/figure>\n\n<\/div>\n<\/div>\n<\/div>\n\n\n\n<p><\/p>\n<\/section>\n<\/div><\/div>\n\n\n\n<p><\/p>\n\n<\/div>\n<\/div>\n<\/div>\n","protected":false},"excerpt":{"rendered":"","protected":false},"author":4,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"template-title-left-aligned.php","meta":{"_acf_changed":false,"footnotes":""},"embl_taxonomy":[],"class_list":["post-3418","page","type-page","status-publish","hentry"],"acf":[],"embl_taxonomy_terms":[],"_links":{"self":[{"href":"https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-json\/wp\/v2\/pages\/3418","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-json\/wp\/v2\/users\/4"}],"replies":[{"embeddable":true,"href":"https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-json\/wp\/v2\/comments?post=3418"}],"version-history":[{"count":18,"href":"https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-json\/wp\/v2\/pages\/3418\/revisions"}],"predecessor-version":[{"id":44783,"href":"https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-json\/wp\/v2\/pages\/3418\/revisions\/44783"}],"wp:attachment":[{"href":"https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-json\/wp\/v2\/media?parent=3418"}],"wp:term":[{"taxonomy":"embl_taxonomy","embeddable":true,"href":"https:\/\/www.embl.org\/about\/info\/imaging-centre\/wp-json\/wp\/v2\/embl_taxonomy?post=3418"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}