{"id":578,"date":"2022-01-01T14:45:16","date_gmt":"2022-01-01T13:45:16","guid":{"rendered":"https:\/\/michelecrevecoeur.ch\/?page_id=578"},"modified":"2022-03-13T15:02:39","modified_gmt":"2022-03-13T14:02:39","slug":"patag-staining","status":"publish","type":"page","link":"https:\/\/michelecrevecoeur.ch\/?page_id=578","title":{"rendered":"PATAg staining of polysaccharides"},"content":{"rendered":"\t\t<div data-elementor-type=\"wp-page\" data-elementor-id=\"578\" class=\"elementor elementor-578\">\n\t\t\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-74ce0454 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"74ce0454\" 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-697b6720\" data-id=\"697b6720\" 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-5b34fb6 elementor-widget elementor-widget-text-editor\" data-id=\"5b34fb6\" 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=\"font-size: medium; color: #003366;\"><strong>Localization of polysaccharides at the ultrastructural level (Transmission Electron Microscope)<br \/>with the\u00a0 PATAg method<\/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;\"><span style=\"color: #003366;\">\u00a0<strong>Principle of PATAg reaction &#8211;\u00a0<\/strong>\u00a0<strong>P<\/strong>eriodic\u00a0<strong>A<\/strong>cid\u00a0<strong>T<\/strong>hiocarbohydrazide\u00a0<strong>A<\/strong>g proteinate<\/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;\"><span style=\"color: #003366;\">The reaction is derived from the PAS staining used in classical histology. It is carried out in 3 main steps. First, a treatment with periodic acid specifically oxidizes vic-glycols(vic for vicinity; glycol for free alcohol group on sugar) into aldehydes. The second step consists in treatment of sections with\u00a0<em>thiocarbohydrazide<\/em>\u00a0(TCH) that condenses on aldehydes groups. Finally TCH was made visible as electron dense particles with silver proteinate.<\/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;\"><strong><span style=\"color: #003366;\">Protocol<\/span><br \/><\/strong>The procedure described here below has been applied to ultrathin sections (80 nm thick) made through organs classically fixed with glutaraldehyde &#8211; OsO<span style=\"position: relative; font-size: 10.5px; line-height: 0; vertical-align: baseline; bottom: -0.25em;\">4\u00a0<\/span>and emdedded in Epon. Sections are collected on gold grids covered with a formvar film. All the steps occur at room temperature.<\/p><ul style=\"margin-bottom: 10px; color: #404040; font-family: arial; font-size: 14px; font-style: normal; font-weight: 400;\"><li>Float section at the surface of 1% periodic acid drops (~ 30 \u00b5l) for 20 min at room temperature. Control grids: deposited on drops of 5% H<span style=\"position: relative; font-size: 10.5px; line-height: 0; vertical-align: baseline; bottom: -0.25em;\">2<\/span>O<span style=\"position: relative; font-size: 10.5px; line-height: 0; vertical-align: baseline; bottom: -0.25em;\">2<\/span>\u00a0instead of periodic acid<\/li><\/ul><ul style=\"margin-bottom: 10px; color: #404040; font-family: arial; font-size: 14px; font-style: normal; font-weight: 400;\" type=\"disc\"><li>Washes 4 x 5 min on drops (~ 50 \u00b5l) of distilled water.<\/li><li>Float the sections on drops (50 \u00b5l) of 0,2 % TCH (Thiocarbohydrazide) in a humid chamber at darkness for 30 min to 72h depending on the material.<\/li><li>Wash on drops of acetic acid of decreasing concentrations: 20 % (4 x 5 min),10% (5 min),5% 5 min, 2% 5 min et 1% 5 min).<\/li><li>Wash with filtered (Millipore) milliQ water 5 x 5 min<\/li><li>Float the sections on drop of aqueous 1% silver &#8211; proteinate for 30 min at room temperature at darkness.<\/li><li>Wash with water 4 x 5 min<\/li><li>The sections are dried then observed with a Transmission Electron Microscope operating at 80 kV.<br \/>Electron microcopes : Philips EM 410;\u00a0FEI TECNAI\u00a0<span style=\"position: relative; font-size: 10.5px; line-height: 0; vertical-align: baseline; top: -0.5em;\">TM<\/span>\u00a0G2 Sphera<\/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;\">The response e.g. dense deposit of silver grains is positive for many kinds of carbohydrates.<\/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;\">Below parts of ultrathin sections (80 nm) made through a primary root of\u00a0<em>Zea mays<\/em>\u00a0and stained with PATAg.<br \/>The first micrograph shows a few cells with postive reaction of starch in amyloplasts (a) and of cell walls that both appear electron dense. N. nucleus<\/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\/7d243f6c56b614ea40b28550c0fbd4ba_f303.jpg\" alt=\"PATAg coiffe racine Zea \" width=\"550\" height=\"443\" \/>.<\/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;\">Below two micrographs with details of positive PATAg reaction in\u00a0<em>Zea mays<\/em>\u00a0root cells<\/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;\">On the right detail of a cell wall separating (p) two cells and of starch grains (ga).\u00a0On the left high magnification view of a starch grain and of the cell wall (paroi) in wich dense deposits of silver appear as small particles.<\/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-b34e237 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"b34e237\" 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-50 elementor-top-column elementor-element elementor-element-4546558\" data-id=\"4546558\" 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-32f8679 elementor-widget elementor-widget-image\" data-id=\"32f8679\" data-element_type=\"widget\" data-e-type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t<img decoding=\"async\" width=\"480\" height=\"407\" src=\"https:\/\/michelecrevecoeur.ch\/wp-content\/uploads\/2022\/01\/9c3a791afa4a601a791c7b26838932ad_f32.jpg\" class=\"attachment-large size-large wp-image-584\" alt=\"\" srcset=\"https:\/\/michelecrevecoeur.ch\/wp-content\/uploads\/2022\/01\/9c3a791afa4a601a791c7b26838932ad_f32.jpg 480w, https:\/\/michelecrevecoeur.ch\/wp-content\/uploads\/2022\/01\/9c3a791afa4a601a791c7b26838932ad_f32-300x254.jpg 300w\" sizes=\"(max-width: 480px) 100vw, 480px\" \/>\t\t\t\t\t\t\t\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<div class=\"elementor-column elementor-col-50 elementor-top-column elementor-element elementor-element-66d6c32\" data-id=\"66d6c32\" 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-fa5f442 elementor-widget elementor-widget-image\" data-id=\"fa5f442\" data-element_type=\"widget\" data-e-type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t<img decoding=\"async\" width=\"500\" height=\"466\" src=\"https:\/\/michelecrevecoeur.ch\/wp-content\/uploads\/2022\/01\/8fc1f3ba2237e6bdff56c6e3b481a1ad_f120.jpg\" class=\"attachment-large size-large wp-image-585\" alt=\"\" srcset=\"https:\/\/michelecrevecoeur.ch\/wp-content\/uploads\/2022\/01\/8fc1f3ba2237e6bdff56c6e3b481a1ad_f120.jpg 500w, https:\/\/michelecrevecoeur.ch\/wp-content\/uploads\/2022\/01\/8fc1f3ba2237e6bdff56c6e3b481a1ad_f120-300x280.jpg 300w\" sizes=\"(max-width: 500px) 100vw, 500px\" \/>\t\t\t\t\t\t\t\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-83429d2 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"83429d2\" 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-cd42f5f\" data-id=\"cd42f5f\" 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-0f36551 elementor-widget elementor-widget-text-editor\" data-id=\"0f36551\" 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=\"text-align: center;\"><span style=\"color: #003366; font-family: arial; font-size: 14px; text-align: center;\">Gallery of electronmicrographs illustrating PATAg reaction.<\/span><\/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-f520850 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"f520850\" 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-857b8e9\" data-id=\"857b8e9\" 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-0d18b18 elementor-widget elementor-widget-image-gallery\" data-id=\"0d18b18\" data-element_type=\"widget\" data-e-type=\"widget\" data-widget_type=\"image-gallery.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<div class=\"elementor-image-gallery\">\n\t\t\t<div id='gallery-1' class='gallery galleryid-578 gallery-columns-3 gallery-size-thumbnail'><figure class='gallery-item'>\n\t\t\t<div class='gallery-icon landscape'>\n\t\t\t\t<a data-elementor-open-lightbox=\"yes\" data-elementor-lightbox-slideshow=\"0d18b18\" data-elementor-lightbox-title=\"74b4db57a53b0d50be196ee7657811ae_f120\" data-e-action-hash=\"#elementor-action%3Aaction%3Dlightbox%26settings%3DeyJpZCI6MzUwMSwidXJsIjoiaHR0cHM6XC9cL21pY2hlbGVjcmV2ZWNvZXVyLmNoXC93cC1jb250ZW50XC91cGxvYWRzXC8yMDIyXC8wM1wvNzRiNGRiNTdhNTNiMGQ1MGJlMTk2ZWU3NjU3ODExYWVfZjEyMC5qcGciLCJzbGlkZXNob3ciOiIwZDE4YjE4In0%3D\" href='https:\/\/michelecrevecoeur.ch\/wp-content\/uploads\/2022\/03\/74b4db57a53b0d50be196ee7657811ae_f120.jpg'><img loading=\"lazy\" decoding=\"async\" width=\"150\" height=\"150\" src=\"https:\/\/michelecrevecoeur.ch\/wp-content\/uploads\/2022\/03\/74b4db57a53b0d50be196ee7657811ae_f120-150x150.jpg\" class=\"attachment-thumbnail size-thumbnail\" alt=\"\" \/><\/a>\n\t\t\t<\/div><\/figure><figure class='gallery-item'>\n\t\t\t<div class='gallery-icon landscape'>\n\t\t\t\t<a data-elementor-open-lightbox=\"yes\" data-elementor-lightbox-slideshow=\"0d18b18\" data-elementor-lightbox-title=\"9512f661751739099e95d1a90754dc61_f303\" data-e-action-hash=\"#elementor-action%3Aaction%3Dlightbox%26settings%3DeyJpZCI6MzUwMiwidXJsIjoiaHR0cHM6XC9cL21pY2hlbGVjcmV2ZWNvZXVyLmNoXC93cC1jb250ZW50XC91cGxvYWRzXC8yMDIyXC8wM1wvOTUxMmY2NjE3NTE3MzkwOTllOTVkMWE5MDc1NGRjNjFfZjMwMy5qcGciLCJzbGlkZXNob3ciOiIwZDE4YjE4In0%3D\" href='https:\/\/michelecrevecoeur.ch\/wp-content\/uploads\/2022\/03\/9512f661751739099e95d1a90754dc61_f303.jpg'><img loading=\"lazy\" decoding=\"async\" width=\"150\" height=\"150\" src=\"https:\/\/michelecrevecoeur.ch\/wp-content\/uploads\/2022\/03\/9512f661751739099e95d1a90754dc61_f303-150x150.jpg\" class=\"attachment-thumbnail size-thumbnail\" alt=\"\" \/><\/a>\n\t\t\t<\/div><\/figure><figure class='gallery-item'>\n\t\t\t<div class='gallery-icon landscape'>\n\t\t\t\t<a data-elementor-open-lightbox=\"yes\" data-elementor-lightbox-slideshow=\"0d18b18\" data-elementor-lightbox-title=\"fa84f5f06fae97700e4cbf770af428bf_f32\" data-e-action-hash=\"#elementor-action%3Aaction%3Dlightbox%26settings%3DeyJpZCI6MzUwMywidXJsIjoiaHR0cHM6XC9cL21pY2hlbGVjcmV2ZWNvZXVyLmNoXC93cC1jb250ZW50XC91cGxvYWRzXC8yMDIyXC8wM1wvZmE4NGY1ZjA2ZmFlOTc3MDBlNGNiZjc3MGFmNDI4YmZfZjMyLmpwZyIsInNsaWRlc2hvdyI6IjBkMThiMTgifQ%3D%3D\" href='https:\/\/michelecrevecoeur.ch\/wp-content\/uploads\/2022\/03\/fa84f5f06fae97700e4cbf770af428bf_f32.jpg'><img loading=\"lazy\" decoding=\"async\" width=\"150\" height=\"150\" src=\"https:\/\/michelecrevecoeur.ch\/wp-content\/uploads\/2022\/03\/fa84f5f06fae97700e4cbf770af428bf_f32-150x150.jpg\" class=\"attachment-thumbnail size-thumbnail\" alt=\"\" \/><\/a>\n\t\t\t<\/div><\/figure>\n\t\t<\/div>\n\t\t<\/div>\n\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>Localization of polysaccharides at the ultrastructural level (Transmission Electron Microscope)with the\u00a0 PATAg method \u00a0Principle of PATAg reaction &#8211;\u00a0\u00a0Periodic\u00a0Acid\u00a0Thiocarbohydrazide\u00a0Ag proteinate The reaction is derived from the PAS staining used in classical histology. It is carried out in 3 main steps. First, a treatment with periodic acid specifically oxidizes vic-glycols(vic for vicinity; glycol for free alcohol group [&hellip;]<\/p>\n","protected":false},"author":2,"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-578","page","type-page","status-publish","hentry"],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v27.2 - https:\/\/yoast.com\/product\/yoast-seo-wordpress\/ -->\n<title>PATAg staining of polysaccharides - Mich\u00e8le Cr\u00e8vecoeur<\/title>\n<meta name=\"description\" content=\"The reaction is derived from the PAS staining used in classical histology. 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