{"id":409,"date":"2021-12-22T16:42:49","date_gmt":"2021-12-22T15:42:49","guid":{"rendered":"https:\/\/michelecrevecoeur.ch\/?page_id=409"},"modified":"2021-12-22T16:53:56","modified_gmt":"2021-12-22T15:53:56","slug":"dapi-staining","status":"publish","type":"page","link":"https:\/\/michelecrevecoeur.ch\/?page_id=409","title":{"rendered":"DAPI staining"},"content":{"rendered":"\t\t
\n\t\t\t\t\t\t
\n\t\t\t\t\t\t
\n\t\t\t\t\t
\n\t\t\t
\n\t\t\t\t\t\t
\n\t\t\t\t
\n\t\t\t\t\t\t\t\t\t

DAPI\u00a0<\/strong>staining of paraffin sections<\/span><\/p>

DAPI (4\u2032,6-diamidino-2-phenylindole) is a fluorochrome that strongly binds with the minor groove of DNA with a preference for the adenine \u2013 thymine base pairs. When bound to double \u2013 stranded DNA, DAPI has an absorption maximum at a wavelength of 358 nm (UV) and its emission maximum at 461 nm (blue). It can also bind to RNA probably at the level of adenine \u2013 uracil clusters. The DAPI\/ RNA complex has an emission maximum at 500 nm a longer wavelength than for the complex DAPI\/DNA and with a lower fluorescence.<\/span><\/p>

Protocol for paraffin section (8\u00a0<\/strong>\u03bc<\/strong>m thick)<\/strong><\/span><\/p>

  • Deparaffinize and hydrate to water.<\/span><\/li>
  • Use a pen to draw a\u00a0hydrophobic<\/em>\u00a0circle\u00a0around<\/em>\u00a0sections<\/span><\/li>
  • Stain with DAPI at appropriate concentration varying from 0.1 mg\/ml to 1 mg\/ml.<\/span><\/li>
  • Add a drop of DAPI on the sections.<\/span><\/li>
  • Wash 3 x 5 min in distilled water<\/span><\/li>
  • Mount in appropriating medium such as Vectashied
    <\/span><\/li>
  • Observe with a fluorescent microscope using appropriate filters.\u00a0
    <\/span>For Inverted Fluorescent Microscope Leica DM IRE 2 the cube filters used contains an excitation filter in UV (340 \u2013 380 nm); a 400 nm dichroic mirror and an emission filter or barrier filter LP 425 nm.<\/span><\/li><\/ul>

    Below : Parts of paraffin sections through a primary\u00a0Zea mays<\/em> root tip<\/strong><\/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

    \n\t\t\t\t\t\t
    \n\t\t\t\t\t
    \n\t\t\t
    \n\t\t\t\t\t\t
    \n\t\t\t\t
    \n\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t\"\"\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
    \n\t\t\t
    \n\t\t\t\t\t\t
    \n\t\t\t\t
    \n\t\t\t\t\t\t\t\t\t\t\t\t
    \n\t\t\t\t\t\t\t\t\t\t\"\"\t\t\t\t\t\t\t\t\t\t\t
    Mitosis (white arrow) are clearly seen.<\/figcaption>\n\t\t\t\t\t\t\t\t\t\t<\/figure>\n\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
    \n\t\t\t\t\t\t
    \n\t\t\t\t\t
    \n\t\t\t
    \n\t\t\t\t\t\t
    \n\t\t\t\t
    \n\t\t\t\t\t\t\t\t\t

    Below : Parts of paraffin sections through root nodules induced by the bacterium strain NGR234\u00a0<\/strong><\/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

    \n\t\t\t\t\t\t
    \n\t\t\t\t\t
    \n\t\t\t
    \n\t\t\t\t\t\t
    \n\t\t\t\t
    \n\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t\"\"\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
    \n\t\t\t
    \n\t\t\t\t\t\t
    \n\t\t\t\t
    \n\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t\"\"\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<\/div>\n\t\t","protected":false},"excerpt":{"rendered":"

    DAPI\u00a0staining of paraffin sections DAPI (4\u2032,6-diamidino-2-phenylindole) is a fluorochrome that strongly binds with the minor groove of DNA with a preference for the adenine \u2013 thymine base pairs. When bound to double \u2013 stranded DNA, DAPI has an absorption maximum at a wavelength of 358 nm (UV) and its emission maximum at 461 nm (blue). […]<\/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-409","page","type-page","status-publish","hentry"],"yoast_head":"\nDAPI 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=409\" \/>\n<meta property=\"og:locale\" content=\"fr_FR\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"DAPI staining - Mich\u00e8le Cr\u00e8vecoeur\" \/>\n<meta property=\"og:description\" content=\"DAPI\u00a0staining of paraffin sections DAPI (4\u2032,6-diamidino-2-phenylindole) is a fluorochrome that strongly binds with the minor groove of DNA with a preference for the adenine \u2013 thymine base pairs. When bound to double \u2013 stranded DNA, DAPI has an absorption maximum at a wavelength of 358 nm (UV) and its emission maximum at 461 nm (blue). […]\" \/>\n<meta property=\"og:url\" content=\"https:\/\/michelecrevecoeur.ch\/?page_id=409\" \/>\n<meta property=\"og:site_name\" content=\"Mich\u00e8le Cr\u00e8vecoeur\" \/>\n<meta property=\"article:modified_time\" content=\"2021-12-22T15:53:56+00:00\" \/>\n<meta property=\"og:image\" content=\"https:\/\/michelecrevecoeur.ch\/wp-content\/uploads\/2021\/12\/DAPI-1.jpg\" \/>\n<meta name=\"twitter:card\" content=\"summary_large_image\" \/>\n<meta name=\"twitter:label1\" content=\"Dur\u00e9e de lecture estim\u00e9e\" \/>\n\t<meta name=\"twitter:data1\" content=\"2 minutes\" \/>\n<script type=\"application\/ld+json\" class=\"yoast-schema-graph\">{\"@context\":\"https:\\\/\\\/schema.org\",\"@graph\":[{\"@type\":\"WebPage\",\"@id\":\"https:\\\/\\\/michelecrevecoeur.ch\\\/?page_id=409\",\"url\":\"https:\\\/\\\/michelecrevecoeur.ch\\\/?page_id=409\",\"name\":\"DAPI staining - Mich\u00e8le Cr\u00e8vecoeur\",\"isPartOf\":{\"@id\":\"https:\\\/\\\/michelecrevecoeur.ch\\\/#website\"},\"primaryImageOfPage\":{\"@id\":\"https:\\\/\\\/michelecrevecoeur.ch\\\/?page_id=409#primaryimage\"},\"image\":{\"@id\":\"https:\\\/\\\/michelecrevecoeur.ch\\\/?page_id=409#primaryimage\"},\"thumbnailUrl\":\"https:\\\/\\\/michelecrevecoeur.ch\\\/wp-content\\\/uploads\\\/2021\\\/12\\\/DAPI-1.jpg\",\"datePublished\":\"2021-12-22T15:42:49+00:00\",\"dateModified\":\"2021-12-22T15:53:56+00:00\",\"breadcrumb\":{\"@id\":\"https:\\\/\\\/michelecrevecoeur.ch\\\/?page_id=409#breadcrumb\"},\"inLanguage\":\"fr-FR\",\"potentialAction\":[{\"@type\":\"ReadAction\",\"target\":[\"https:\\\/\\\/michelecrevecoeur.ch\\\/?page_id=409\"]}]},{\"@type\":\"ImageObject\",\"inLanguage\":\"fr-FR\",\"@id\":\"https:\\\/\\\/michelecrevecoeur.ch\\\/?page_id=409#primaryimage\",\"url\":\"https:\\\/\\\/michelecrevecoeur.ch\\\/wp-content\\\/uploads\\\/2021\\\/12\\\/DAPI-1.jpg\",\"contentUrl\":\"https:\\\/\\\/michelecrevecoeur.ch\\\/wp-content\\\/uploads\\\/2021\\\/12\\\/DAPI-1.jpg\",\"width\":649,\"height\":488},{\"@type\":\"BreadcrumbList\",\"@id\":\"https:\\\/\\\/michelecrevecoeur.ch\\\/?page_id=409#breadcrumb\",\"itemListElement\":[{\"@type\":\"ListItem\",\"position\":1,\"name\":\"Accueil\",\"item\":\"https:\\\/\\\/michelecrevecoeur.ch\\\/\"},{\"@type\":\"ListItem\",\"position\":2,\"name\":\"DAPI staining\"}]},{\"@type\":\"WebSite\",\"@id\":\"https:\\\/\\\/michelecrevecoeur.ch\\\/#website\",\"url\":\"https:\\\/\\\/michelecrevecoeur.ch\\\/\",\"name\":\"Mich\u00e8le Cr\u00e8vecoeur\",\"description\":\"Plant Microscopy\",\"publisher\":{\"@id\":\"https:\\\/\\\/michelecrevecoeur.ch\\\/#\\\/schema\\\/person\\\/dfdf9ef4c6d746dfdecdceb6e2986d66\"},\"potentialAction\":[{\"@type\":\"SearchAction\",\"target\":{\"@type\":\"EntryPoint\",\"urlTemplate\":\"https:\\\/\\\/michelecrevecoeur.ch\\\/?s={search_term_string}\"},\"query-input\":{\"@type\":\"PropertyValueSpecification\",\"valueRequired\":true,\"valueName\":\"search_term_string\"}}],\"inLanguage\":\"fr-FR\"},{\"@type\":[\"Person\",\"Organization\"],\"@id\":\"https:\\\/\\\/michelecrevecoeur.ch\\\/#\\\/schema\\\/person\\\/dfdf9ef4c6d746dfdecdceb6e2986d66\",\"name\":\"Mich\u00e8le\",\"image\":{\"@type\":\"ImageObject\",\"inLanguage\":\"fr-FR\",\"@id\":\"https:\\\/\\\/secure.gravatar.com\\\/avatar\\\/5ad72d9990ca501b7348a22de838f81a21f31163f3372e910adcb88b005c4cca?s=96&d=mm&r=g\",\"url\":\"https:\\\/\\\/secure.gravatar.com\\\/avatar\\\/5ad72d9990ca501b7348a22de838f81a21f31163f3372e910adcb88b005c4cca?s=96&d=mm&r=g\",\"contentUrl\":\"https:\\\/\\\/secure.gravatar.com\\\/avatar\\\/5ad72d9990ca501b7348a22de838f81a21f31163f3372e910adcb88b005c4cca?s=96&d=mm&r=g\",\"caption\":\"Mich\u00e8le\"},\"logo\":{\"@id\":\"https:\\\/\\\/secure.gravatar.com\\\/avatar\\\/5ad72d9990ca501b7348a22de838f81a21f31163f3372e910adcb88b005c4cca?s=96&d=mm&r=g\"},\"sameAs\":[\"http:\\\/\\\/michelecrevecoeur.ch\"]}]}<\/script>\n<!-- \/ Yoast SEO plugin. -->","yoast_head_json":{"title":"DAPI staining - Mich\u00e8le Cr\u00e8vecoeur","robots":{"index":"index","follow":"follow","max-snippet":"max-snippet:-1","max-image-preview":"max-image-preview:large","max-video-preview":"max-video-preview:-1"},"canonical":"https:\/\/michelecrevecoeur.ch\/?page_id=409","og_locale":"fr_FR","og_type":"article","og_title":"DAPI staining - Mich\u00e8le Cr\u00e8vecoeur","og_description":"DAPI\u00a0staining of paraffin sections DAPI (4\u2032,6-diamidino-2-phenylindole) is a fluorochrome that strongly binds with the minor groove of DNA with a preference for the adenine \u2013 thymine base pairs. When bound to double \u2013 stranded DNA, DAPI has an absorption maximum at a wavelength of 358 nm (UV) and its emission maximum at 461 nm (blue). […]","og_url":"https:\/\/michelecrevecoeur.ch\/?page_id=409","og_site_name":"Mich\u00e8le Cr\u00e8vecoeur","article_modified_time":"2021-12-22T15:53:56+00:00","og_image":[{"url":"https:\/\/michelecrevecoeur.ch\/wp-content\/uploads\/2021\/12\/DAPI-1.jpg","type":"","width":"","height":""}],"twitter_card":"summary_large_image","twitter_misc":{"Dur\u00e9e de lecture estim\u00e9e":"2 minutes"},"schema":{"@context":"https:\/\/schema.org","@graph":[{"@type":"WebPage","@id":"https:\/\/michelecrevecoeur.ch\/?page_id=409","url":"https:\/\/michelecrevecoeur.ch\/?page_id=409","name":"DAPI staining - Mich\u00e8le Cr\u00e8vecoeur","isPartOf":{"@id":"https:\/\/michelecrevecoeur.ch\/#website"},"primaryImageOfPage":{"@id":"https:\/\/michelecrevecoeur.ch\/?page_id=409#primaryimage"},"image":{"@id":"https:\/\/michelecrevecoeur.ch\/?page_id=409#primaryimage"},"thumbnailUrl":"https:\/\/michelecrevecoeur.ch\/wp-content\/uploads\/2021\/12\/DAPI-1.jpg","datePublished":"2021-12-22T15:42:49+00:00","dateModified":"2021-12-22T15:53:56+00:00","breadcrumb":{"@id":"https:\/\/michelecrevecoeur.ch\/?page_id=409#breadcrumb"},"inLanguage":"fr-FR","potentialAction":[{"@type":"ReadAction","target":["https:\/\/michelecrevecoeur.ch\/?page_id=409"]}]},{"@type":"ImageObject","inLanguage":"fr-FR","@id":"https:\/\/michelecrevecoeur.ch\/?page_id=409#primaryimage","url":"https:\/\/michelecrevecoeur.ch\/wp-content\/uploads\/2021\/12\/DAPI-1.jpg","contentUrl":"https:\/\/michelecrevecoeur.ch\/wp-content\/uploads\/2021\/12\/DAPI-1.jpg","width":649,"height":488},{"@type":"BreadcrumbList","@id":"https:\/\/michelecrevecoeur.ch\/?page_id=409#breadcrumb","itemListElement":[{"@type":"ListItem","position":1,"name":"Accueil","item":"https:\/\/michelecrevecoeur.ch\/"},{"@type":"ListItem","position":2,"name":"DAPI staining"}]},{"@type":"WebSite","@id":"https:\/\/michelecrevecoeur.ch\/#website","url":"https:\/\/michelecrevecoeur.ch\/","name":"Mich\u00e8le Cr\u00e8vecoeur","description":"Plant Microscopy","publisher":{"@id":"https:\/\/michelecrevecoeur.ch\/#\/schema\/person\/dfdf9ef4c6d746dfdecdceb6e2986d66"},"potentialAction":[{"@type":"SearchAction","target":{"@type":"EntryPoint","urlTemplate":"https:\/\/michelecrevecoeur.ch\/?s={search_term_string}"},"query-input":{"@type":"PropertyValueSpecification","valueRequired":true,"valueName":"search_term_string"}}],"inLanguage":"fr-FR"},{"@type":["Person","Organization"],"@id":"https:\/\/michelecrevecoeur.ch\/#\/schema\/person\/dfdf9ef4c6d746dfdecdceb6e2986d66","name":"Mich\u00e8le","image":{"@type":"ImageObject","inLanguage":"fr-FR","@id":"https:\/\/secure.gravatar.com\/avatar\/5ad72d9990ca501b7348a22de838f81a21f31163f3372e910adcb88b005c4cca?s=96&d=mm&r=g","url":"https:\/\/secure.gravatar.com\/avatar\/5ad72d9990ca501b7348a22de838f81a21f31163f3372e910adcb88b005c4cca?s=96&d=mm&r=g","contentUrl":"https:\/\/secure.gravatar.com\/avatar\/5ad72d9990ca501b7348a22de838f81a21f31163f3372e910adcb88b005c4cca?s=96&d=mm&r=g","caption":"Mich\u00e8le"},"logo":{"@id":"https:\/\/secure.gravatar.com\/avatar\/5ad72d9990ca501b7348a22de838f81a21f31163f3372e910adcb88b005c4cca?s=96&d=mm&r=g"},"sameAs":["http:\/\/michelecrevecoeur.ch"]}]}},"_links":{"self":[{"href":"https:\/\/michelecrevecoeur.ch\/index.php?rest_route=\/wp\/v2\/pages\/409","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/michelecrevecoeur.ch\/index.php?rest_route=\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/michelecrevecoeur.ch\/index.php?rest_route=\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/michelecrevecoeur.ch\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/michelecrevecoeur.ch\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=409"}],"version-history":[{"count":7,"href":"https:\/\/michelecrevecoeur.ch\/index.php?rest_route=\/wp\/v2\/pages\/409\/revisions"}],"predecessor-version":[{"id":436,"href":"https:\/\/michelecrevecoeur.ch\/index.php?rest_route=\/wp\/v2\/pages\/409\/revisions\/436"}],"wp:attachment":[{"href":"https:\/\/michelecrevecoeur.ch\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=409"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}