{"id":426,"date":"2021-12-22T16:50:29","date_gmt":"2021-12-22T15:50:29","guid":{"rendered":"https:\/\/michelecrevecoeur.ch\/?page_id=426"},"modified":"2021-12-22T16:51:53","modified_gmt":"2021-12-22T15:51:53","slug":"calcofluor-white-staining","status":"publish","type":"page","link":"https:\/\/michelecrevecoeur.ch\/?page_id=426","title":{"rendered":"Calcofluor White staining"},"content":{"rendered":"\t\t<div data-elementor-type=\"wp-page\" data-elementor-id=\"426\" class=\"elementor elementor-426\">\n\t\t\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-6f1c7eb elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"6f1c7eb\" data-element_type=\"section\" data-e-type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-2bddea78\" data-id=\"2bddea78\" data-element_type=\"column\" data-e-type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t<div class=\"elementor-element elementor-element-58638cd4 elementor-widget elementor-widget-text-editor\" data-id=\"58638cd4\" data-element_type=\"widget\" data-e-type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t<p style=\"margin-top: 15px; margin-bottom: 15px; line-height: 25.2px; color: #404040; font-family: arial; font-size: 14px; font-style: normal; font-weight: 400; text-align: center;\"><span style=\"color: #003366; font-size: medium;\"><strong>Calcofluor White staining<\/strong><\/span><\/p><p style=\"margin-top: 15px; margin-bottom: 15px; line-height: 25.2px; color: #404040; font-family: arial; font-size: 14px; font-style: normal; font-weight: 400;\">Calcofluor White\u00a0<span lang=\"EN-US\">(Fluorescent Brightner 28)\u00a0<\/span>is a fluorescent blue dye that binds to polysaccharides. This reagent can bind via hydrogen bonds to \u03b2 (1\u2192 4) et \u03b2 (1\u21923) polysaccharides. It shows a strong affinity for cellulose forming hydrogen bonds with free hydroxyls groups. It is useful to see all cell walls in plant materials using fluorescence microscopy with UV excitation at 365 nm and emission at ~ 440 nm. In medicine it is used for detection of fungal pathogens as it binds to chitin.<\/p><p style=\"margin-top: 15px; margin-bottom: 15px; line-height: 25.2px; color: #404040; font-family: arial; font-size: 14px; font-style: normal; font-weight: 400;\"><span style=\"color: #003366;\"><strong>Protocol for paraffin sections.<\/strong><\/span>\u00a0<\/p><ul style=\"margin-bottom: 10px; color: #404040; font-family: arial; font-size: 14px; font-style: normal; font-weight: 400;\"><li>The protocol is appropriated for plant organs fixed with FAA or aldehydes and embedded in paraffin.<\/li><li>Deparaffinize and hydrate to water<\/li><li>Use a pen to draw a\u00a0<em>hydrophobic<\/em>\u00a0circle\u00a0<em>around<\/em>\u00a0sections.<\/li><li>Carefully remove the excess of water and add a drop (100 \u00b5l) of aqueous solution of Calcofluor (Sigma F3543) at a concentration varying from 0.01 to 0,005%. Control of autofluorescence: sections with a drop of distilled water.<\/li><li>\u00a0Stain at darkness for 30 sec to 15 min. The concentration and length of staining varies according to the material and the fixative. Examples: for\u00a0<em>Zea mays<\/em>\u00a0roots fixed with FAA 30 sec in 0,01%; for\u00a0<em>Quercus<\/em>\u00a0roots fixed with formaldehyde \u2013 glutaraldehyde, 5 min with 0,1%.<\/li><li>Wash slides thoroughly with distilled water with gentle agitation.<\/li><li>Mount in appropriating medium such as Vectashied. (Antifading mounting medium)<\/li><li>Observation: fluorescence microscope with appropriated filters. For Leica DM IRE 2: A filter for excitation (UV: 340 \u2013 380 nm); For Confocal Leica SP2: excitation with laser 405 nm; emission: 450 to 547 nm.<\/li><\/ul><p style=\"margin-top: 15px; margin-bottom: 15px; line-height: 25.2px; color: #404040; font-family: arial; font-size: 14px; font-style: normal; font-weight: 400;\">\u00a0A stock solution of calcofluor can be prepared and stored at darkness at 4\u00b0C for months.<\/p><p style=\"margin-top: 15px; margin-bottom: 15px; line-height: 25.2px; color: #404040; font-family: arial; font-size: 14px; font-style: normal; font-weight: 400;\">Two micrographs of paraffin sections stained with calcofluor white are shown\u00a0below. They were observed and imaged with a Leica DM IRE2 microscope (excitation at 340-380 nm; emission at 425 nm) equipped with a Leica DC300F camera.<\/p><p style=\"margin-top: 15px; margin-bottom: 15px; line-height: 25.2px; color: #404040; font-family: arial; font-size: 14px; font-style: normal; font-weight: 400;\">The fisrt one is a cross section through a\u00a0<em>Zea mays<\/em>\u00a0primary root with root cap at the outside. Cellulosic cell walls are all fluorescent and there is autofluorescence of lignin and suberin. cc : central cylinder.<\/p><p style=\"margin-top: 15px; margin-bottom: 15px; line-height: 25.2px; color: #404040; font-family: arial; font-size: 14px; font-style: normal; font-weight: 400;\"><img fetchpriority=\"high\" decoding=\"async\" style=\"border-width: 0px; border-style: initial; display: block; margin-left: auto; margin-right: auto;\" src=\"https:\/\/www.unige.ch\/sciences\/biologie\/bioveg\/crevecoeur\/files\/cache\/0f754458e920610d1b987d65aefc8127_f512.jpg\" alt=\"Racine Zea calcofluor\" width=\"700\" height=\"633\" \/><\/p>\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-0a11e12 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"0a11e12\" data-element_type=\"section\" data-e-type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-c2afc3f\" data-id=\"c2afc3f\" data-element_type=\"column\" data-e-type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t<div class=\"elementor-element elementor-element-2945b13 elementor-widget elementor-widget-text-editor\" data-id=\"2945b13\" data-element_type=\"widget\" data-e-type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t<p style=\"margin-top: 15px; margin-bottom: 15px; line-height: 25.2px; color: #404040; font-family: arial; font-size: 14px; font-style: normal; font-weight: 400; text-align: center;\"><strong>Microphotograph of a paraffin section through an\u00a0<em>Arabidopsis thaliana\u00a0<\/em>embryo<\/strong><\/p><p style=\"margin-top: 15px; margin-bottom: 15px; line-height: 25.2px; color: #404040; font-family: arial; font-size: 14px; font-style: normal; font-weight: 400;\"><img decoding=\"async\" style=\"border-width: 0px; border-style: initial; display: block; margin-left: auto; margin-right: auto;\" src=\"https:\/\/www.unige.ch\/sciences\/biologie\/bioveg\/crevecoeur\/files\/cache\/f9cf23863cc0eef6c088aa8ea29035fd_f513.jpg\" alt=\"Embryon Arabidopsis calcofluor\" width=\"700\" height=\"560\" \/><\/p>\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<\/div>\n\t\t","protected":false},"excerpt":{"rendered":"<p>Calcofluor White staining Calcofluor White\u00a0(Fluorescent Brightner 28)\u00a0is a fluorescent blue dye that binds to polysaccharides. This reagent can bind via hydrogen bonds to \u03b2 (1\u2192 4) et \u03b2 (1\u21923) polysaccharides. It shows a strong affinity for cellulose forming hydrogen bonds with free hydroxyls groups. It is useful to see all cell walls in plant materials [&hellip;]<\/p>\n","protected":false},"author":1,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"site-sidebar-layout":"no-sidebar","site-content-layout":"page-builder","ast-site-content-layout":"default","site-content-style":"default","site-sidebar-style":"default","ast-global-header-display":"","ast-banner-title-visibility":"","ast-main-header-display":"","ast-hfb-above-header-display":"","ast-hfb-below-header-display":"","ast-hfb-mobile-header-display":"","site-post-title":"disabled","ast-breadcrumbs-content":"","ast-featured-img":"disabled","footer-sml-layout":"","ast-disable-related-posts":"","theme-transparent-header-meta":"","adv-header-id-meta":"","stick-header-meta":"","header-above-stick-meta":"","header-main-stick-meta":"","header-below-stick-meta":"","astra-migrate-meta-layouts":"default","ast-page-background-enabled":"default","ast-page-background-meta":{"desktop":{"background-color":"var(--ast-global-color-4)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"tablet":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"mobile":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""}},"ast-content-background-meta":{"desktop":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"tablet":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"mobile":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""}},"footnotes":""},"class_list":["post-426","page","type-page","status-publish","hentry"],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v27.8 - https:\/\/yoast.com\/product\/yoast-seo-wordpress\/ -->\n<title>Calcofluor White staining - Mich\u00e8le Cr\u00e8vecoeur<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/michelecrevecoeur.ch\/?page_id=426\" \/>\n<meta property=\"og:locale\" content=\"fr_FR\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Calcofluor White staining - Mich\u00e8le Cr\u00e8vecoeur\" \/>\n<meta property=\"og:description\" content=\"Calcofluor White staining Calcofluor White\u00a0(Fluorescent Brightner 28)\u00a0is a fluorescent blue dye that binds to polysaccharides. 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