{"id":785,"date":"2017-08-10T15:08:46","date_gmt":"2017-08-10T15:08:46","guid":{"rendered":"http:\/\/physioweb.uvm.edu\/warshaw-lab\/?page_id=785"},"modified":"2017-08-10T15:08:46","modified_gmt":"2017-08-10T15:08:46","slug":"storm-microscopy","status":"publish","type":"page","link":"https:\/\/physioweb.uvm.edu\/warshaw-lab\/storm-microscopy\/","title":{"rendered":"STORM Microscopy"},"content":{"rendered":"<table style=\"width: 900px;\" border=\"0\" cellspacing=\"5\" cellpadding=\"5\">\n<tbody>\n<tr>\n<td width=\"25%\"><div id='Techniques_Menu' class='widgets_on_page wop_tiny1  wop_small1  wop_medium1  wop_large1  wop_wide1'>\n\t\t\t<ul><li id=\"nav_menu-8\" class=\"widget widget_nav_menu\"><div class=\"menu-techniques-container\"><ul id=\"menu-techniques\" class=\"menu\"><li id=\"menu-item-322\" class=\"menu-item menu-item-type-post_type menu-item-object-page menu-item-322\"><a href=\"https:\/\/physioweb.uvm.edu\/warshaw-lab\/techniques\/in-vitro-motility-assay\/\">In Vitro Motility Assay<\/a><\/li>\n<li id=\"menu-item-324\" class=\"menu-item menu-item-type-post_type menu-item-object-page menu-item-324\"><a href=\"https:\/\/physioweb.uvm.edu\/warshaw-lab\/techniques\/laser-trap-assay\/\">Laser Trap Assay<\/a><\/li>\n<li id=\"menu-item-326\" class=\"menu-item menu-item-type-post_type menu-item-object-page menu-item-326\"><a href=\"https:\/\/physioweb.uvm.edu\/warshaw-lab\/techniques\/tirfm-laser-trap-assay\/\">TIRFM \u2013 Laser Trap Assay<\/a><\/li>\n<li id=\"menu-item-800\" class=\"menu-item menu-item-type-post_type menu-item-object-page menu-item-800\"><a href=\"https:\/\/physioweb.uvm.edu\/warshaw-lab\/storm-microscopy\/\">STORM Microscopy<\/a><\/li>\n<\/ul><\/div><\/li><\/ul><\/div><!-- widgets_on_page --><\/td>\n<td width=\"623\">\n<p><strong>Super-Resolution STORM Microscopy<\/strong><\/p>\n<p><strong>ST<\/strong>ochastic <strong>O<\/strong>ptical <strong>R<\/strong>econstruction <strong>M<\/strong>icroscopy (STORM) uses the ability to identify single <a href=\"http:\/\/physioweb.uvm.edu\/warshaw-lab\/wp-content\/uploads\/sites\/2\/2017\/08\/Tirf-STORM-Image.png\"><img loading=\"lazy\" class=\"alignright size-medium wp-image-790\" src=\"http:\/\/physioweb.uvm.edu\/warshaw-lab\/wp-content\/uploads\/sites\/2\/2017\/08\/Tirf-STORM-Image-277x300.png\" alt=\"\" width=\"277\" height=\"300\" srcset=\"https:\/\/physioweb.uvm.edu\/warshaw-lab\/wp-content\/uploads\/sites\/2\/2017\/08\/Tirf-STORM-Image-277x300.png 277w, https:\/\/physioweb.uvm.edu\/warshaw-lab\/wp-content\/uploads\/sites\/2\/2017\/08\/Tirf-STORM-Image-768x832.png 768w, https:\/\/physioweb.uvm.edu\/warshaw-lab\/wp-content\/uploads\/sites\/2\/2017\/08\/Tirf-STORM-Image-945x1024.png 945w, https:\/\/physioweb.uvm.edu\/warshaw-lab\/wp-content\/uploads\/sites\/2\/2017\/08\/Tirf-STORM-Image.png 1038w\" sizes=\"(max-width: 277px) 100vw, 277px\" \/><\/a>fluorophores with ~20nm spatial resolution in order to effectively improve the resolution of the light microscope 10-fold. As an example, compare the STORM and TIRF images of the same field of actin filaments, labeled with fluorescent phalloidin and adhered to a glass surface. To reconstruct the super-resolution image, fluorophores are forced into the dark state from which they recover stochastically and fluoresce. As they fluoresce, their positions (i.e. center of point spread functions) are identified with sub-pixel resolution and the positions for all fluorophores on a single actin filament are then used to reconstruct the super-resolution image.<\/p>\n<p>&nbsp;<\/p>\n<p>&nbsp;<\/p>\n<p>By placing a cylindrical lens in the optical path, an optically-induced astigmatism can be used to <a href=\"http:\/\/physioweb.uvm.edu\/warshaw-lab\/wp-content\/uploads\/sites\/2\/2017\/08\/STORM-Actin-Mesh.png\"><img loading=\"lazy\" class=\"alignright size-medium wp-image-793\" src=\"http:\/\/physioweb.uvm.edu\/warshaw-lab\/wp-content\/uploads\/sites\/2\/2017\/08\/STORM-Actin-Mesh-300x300.png\" alt=\"\" width=\"300\" height=\"300\" srcset=\"https:\/\/physioweb.uvm.edu\/warshaw-lab\/wp-content\/uploads\/sites\/2\/2017\/08\/STORM-Actin-Mesh-300x300.png 300w, https:\/\/physioweb.uvm.edu\/warshaw-lab\/wp-content\/uploads\/sites\/2\/2017\/08\/STORM-Actin-Mesh-150x150.png 150w, https:\/\/physioweb.uvm.edu\/warshaw-lab\/wp-content\/uploads\/sites\/2\/2017\/08\/STORM-Actin-Mesh-768x768.png 768w, https:\/\/physioweb.uvm.edu\/warshaw-lab\/wp-content\/uploads\/sites\/2\/2017\/08\/STORM-Actin-Mesh-1024x1024.png 1024w, https:\/\/physioweb.uvm.edu\/warshaw-lab\/wp-content\/uploads\/sites\/2\/2017\/08\/STORM-Actin-Mesh.png 1125w\" sizes=\"(max-width: 300px) 100vw, 300px\" \/><\/a>resolve the Z-position of a fluorophore within the focal plane. As an example, the height of fluorescently-labeled actin filaments suspended from 3\u00b5m beads can be resolved with 50nm resolution.<\/p>\n<p>&nbsp;<\/p>\n<p>&nbsp;<\/p>\n<p>&nbsp;<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p><\/p>\n","protected":false},"excerpt":{"rendered":"<p>Super-Resolution STORM Microscopy STochastic Optical Reconstruction Microscopy (STORM) uses the ability to identify single fluorophores with ~20nm spatial resolution in order to effectively improve the resolution of the light microscope 10-fold. As an example, compare the STORM and TIRF images &hellip; <a href=\"https:\/\/physioweb.uvm.edu\/warshaw-lab\/storm-microscopy\/\">Continue reading <span class=\"meta-nav\">&rarr;<\/span><\/a><\/p>\n","protected":false},"author":5,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":[],"_links":{"self":[{"href":"https:\/\/physioweb.uvm.edu\/warshaw-lab\/wp-json\/wp\/v2\/pages\/785"}],"collection":[{"href":"https:\/\/physioweb.uvm.edu\/warshaw-lab\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/physioweb.uvm.edu\/warshaw-lab\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/physioweb.uvm.edu\/warshaw-lab\/wp-json\/wp\/v2\/users\/5"}],"replies":[{"embeddable":true,"href":"https:\/\/physioweb.uvm.edu\/warshaw-lab\/wp-json\/wp\/v2\/comments?post=785"}],"version-history":[{"count":12,"href":"https:\/\/physioweb.uvm.edu\/warshaw-lab\/wp-json\/wp\/v2\/pages\/785\/revisions"}],"predecessor-version":[{"id":799,"href":"https:\/\/physioweb.uvm.edu\/warshaw-lab\/wp-json\/wp\/v2\/pages\/785\/revisions\/799"}],"wp:attachment":[{"href":"https:\/\/physioweb.uvm.edu\/warshaw-lab\/wp-json\/wp\/v2\/media?parent=785"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}