{"id":634,"date":"2015-02-09T15:44:51","date_gmt":"2015-02-09T15:44:51","guid":{"rendered":"http:\/\/joekitz.com\/LungMap\/?p=634"},"modified":"2018-08-17T02:49:53","modified_gmt":"2018-08-17T02:49:53","slug":"e16-5_c57bl6_calca_scgba1a_acta2_immunofluorescence","status":"publish","type":"post","link":"https:\/\/research.cchmc.org\/lungimage\/?p=634","title":{"rendered":"E16.5 C57BL6 SCGB1A1, CALCA, and ACTA2 Confocal Imaging"},"content":{"rendered":"\t\t<div data-elementor-type=\"wp-post\" data-elementor-id=\"634\" class=\"elementor elementor-634 elementor-bc-flex-widget\" data-elementor-post-type=\"post\">\n\t\t\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-519c089f elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"519c089f\" data-element_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-78aee63f\" data-id=\"78aee63f\" data-element_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-33136461 elementor-widget elementor-widget-theme-post-title elementor-page-title elementor-widget-heading\" data-id=\"33136461\" data-element_type=\"widget\" data-widget_type=\"theme-post-title.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<h1 class=\"elementor-heading-title elementor-size-large\">E16.5 C57BL6 SCGB1A1, CALCA, and ACTA2 Confocal Imaging<\/h1>\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<section class=\"elementor-section elementor-inner-section elementor-element elementor-element-2290dce5 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"2290dce5\" data-element_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-inner-column elementor-element elementor-element-49a0096a\" data-id=\"49a0096a\" data-element_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-235964cc elementor-widget elementor-widget-shortcode\" data-id=\"235964cc\" data-element_type=\"widget\" data-widget_type=\"shortcode.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<div class=\"elementor-shortcode\"><div class=\"wonderplugingallery-container\" id=\"wonderplugingallery-container-8\" style=\"max-width:550px;margin:0 auto;\"><div class=\"wonderplugingallery\" id=\"wonderplugingallery-8\" data-galleryid=\"8\" data-width=\"550\" data-height=\"550\" data-skin=\"light\" data-autoslide=\"false\" data-autoplayvideo=\"false\" data-html5player=\"true\" data-responsive=\"true\" data-fullwidth=\"false\" data-showtitle=\"false\" data-showdescription=\"false\" data-showplaybutton=\"false\" data-showfullscreenbutton=\"true\" data-showtimer=\"false\" data-showcarousel=\"true\" data-galleryshadow=\"true\" data-slideshadow=\"true\" data-thumbshowtitle=\"false\" data-thumbshadow=\"true\" data-lightboxshowtitle=\"true\" data-lightboxshowdescription=\"true\" data-specifyid=\"true\" data-showimgtitle=\"false\" data-titlesmallscreen=\"false\" data-initsocial=\"false\" data-showsocial=\"false\" data-showemail=\"false\" data-showfacebook=\"false\" data-showtwitter=\"false\" data-showpinterest=\"false\" data-socialrotateeffect=\"false\" data-doshortcodeontext=\"false\" data-googleanalyticsaccount=\"\" data-resizemode=\"fit\" data-imagetoolboxmode=\"show\" data-effect=\"fade\" data-padding=\"12\" data-bgcolor=\"\" data-bgimage=\"\" data-thumbwidth=\"100\" data-thumbheight=\"100\" data-thumbgap=\"8\" data-thumbrowgap=\"16\" data-lightboxtextheight=\"72\" data-lightboxtitlecss=\"{color:#333333; font:bold 12px Arial,Helvetica,sans-serif; overflow:hidden; white-space:normal; text-align:Center; line-height:18px;}\" data-lightboxdescriptioncss=\"{color:#333333; font:normal 12px Arial,Helvetica,sans-serif; overflow:hidden; white-space:normal; text-align:Center; line-height:14px;}\" data-titlecss=\"{color:#ffffff; font-size:14px; font-family:Armata, sans-serif, Arial; overflow:hidden; white-space:normal; text-align:Center; padding:10px 0px 10px 10px;  background:rgb(102, 102, 102) transparent; background:rgba(102, 102, 102, 0.6);}\" data-descriptioncss=\"{color:#ffffff; font-size:12px; font-family:Armata, sans-serif, Arial; overflow:hidden; white-space:normal; text-align:Center; padding:0px 0px 10px 10px;  background:rgb(102, 102, 102) transparent; background:rgba(102, 102, 102, 0.6);}\" data-jsfolder=\"https:\/\/research.cchmc.org\/lungimage\/wp-content\/plugins\/wonderplugin-gallery\/engine\/\" style=\"display:none;\" ><a class=\"html5galleryimglink\" href=\"https:\/\/research.cchmc.org\/lungimage\/wp-content\/uploads\/2015\/04\/2014-004-014_C57Bl6_E16.5_LMM.14.24.4.53_SCGA1A1_CALCA_ACTA2-01.jpg\" data-mediatype=1><img decoding=\"async\" class=\"lazyload html5galleryimg html5gallery-tn-image\" src=\"data:image\/svg+xml,%3Csvg%20xmlns%3D%27http%3A%2F%2Fwww.w3.org%2F2000%2Fsvg%27%20width%3D%271024%27%20height%3D%271024%27%20viewBox%3D%270%200%201024%201024%27%3E%3Crect%20width%3D%271024%27%20height%3D%271024%27%20fill-opacity%3D%220%22%2F%3E%3C%2Fsvg%3E\" data-orig-src=\"https:\/\/research.cchmc.org\/lungimage\/wp-content\/uploads\/2015\/04\/2014-004-014_C57Bl6_E16.5_LMM.14.24.4.53_SCGA1A1_CALCA_ACTA2-01.jpg\" alt=\"E16.5 Immunofluorescence and Confocal Imaging Showing Expression of CALCA, SCGB1A1, and ACTA2\" data-description=\"20X Tile Scan\"><\/a><div class=\"html5gallery-info\"><div class=\"html5gallery-title\">E16.5 Immunofluorescence and Confocal Imaging Showing Expression of CALCA, SCGB1A1, and ACTA2<\/div><div class=\"html5gallery-description\">20X Tile Scan<\/div><\/div><a class=\"html5galleryimglink\" href=\"https:\/\/research.cchmc.org\/lungimage\/wp-content\/uploads\/2015\/04\/2014-004-014_C57Bl6_E16.5_LMM.14.24.4.53_SCGA1A1_CALCA_ACTA2-02.jpg\" data-mediatype=1><img decoding=\"async\" class=\"lazyload html5galleryimg html5gallery-tn-image\" src=\"data:image\/svg+xml,%3Csvg%20xmlns%3D%27http%3A%2F%2Fwww.w3.org%2F2000%2Fsvg%27%20width%3D%271024%27%20height%3D%271024%27%20viewBox%3D%270%200%201024%201024%27%3E%3Crect%20width%3D%271024%27%20height%3D%271024%27%20fill-opacity%3D%220%22%2F%3E%3C%2Fsvg%3E\" data-orig-src=\"https:\/\/research.cchmc.org\/lungimage\/wp-content\/uploads\/2015\/04\/2014-004-014_C57Bl6_E16.5_LMM.14.24.4.53_SCGA1A1_CALCA_ACTA2-02.jpg\" alt=\"E16.5 Immunofluorescence and Confocal Imaging Showing Expression of CALCA, SCGB1A1, and ACTA2\" data-description=\"20X\"><\/a><div class=\"html5gallery-info\"><div class=\"html5gallery-title\">E16.5 Immunofluorescence and Confocal Imaging Showing Expression of CALCA, SCGB1A1, and ACTA2<\/div><div class=\"html5gallery-description\">20X<\/div><\/div><a class=\"html5galleryimglink\" href=\"https:\/\/research.cchmc.org\/lungimage\/wp-content\/uploads\/2015\/04\/2014-004-014_C57Bl6_E16.5_LMM.14.24.4.53_SCGA1A1_CALCA_ACTA2-03.jpg\" data-mediatype=1><img decoding=\"async\" class=\"lazyload html5galleryimg html5gallery-tn-image\" src=\"data:image\/svg+xml,%3Csvg%20xmlns%3D%27http%3A%2F%2Fwww.w3.org%2F2000%2Fsvg%27%20width%3D%271024%27%20height%3D%271024%27%20viewBox%3D%270%200%201024%201024%27%3E%3Crect%20width%3D%271024%27%20height%3D%271024%27%20fill-opacity%3D%220%22%2F%3E%3C%2Fsvg%3E\" data-orig-src=\"https:\/\/research.cchmc.org\/lungimage\/wp-content\/uploads\/2015\/04\/2014-004-014_C57Bl6_E16.5_LMM.14.24.4.53_SCGA1A1_CALCA_ACTA2-03.jpg\" alt=\"E16.5 Immunofluorescence and Confocal Imaging Showing Expression of CALCA, SCGB1A1, and ACTA2\" data-description=\"60X\"><\/a><div class=\"html5gallery-info\"><div class=\"html5gallery-title\">E16.5 Immunofluorescence and Confocal Imaging Showing Expression of CALCA, SCGB1A1, and ACTA2<\/div><div class=\"html5gallery-description\">60X<\/div><\/div><a class=\"html5galleryimglink\" href=\"https:\/\/research.cchmc.org\/lungimage\/wp-content\/uploads\/2015\/04\/2014-004-014_C57Bl6_E16.5_LMM.14.24.4.53_SCGA1A1_CALCA_ACTA2-04.jpg\" data-mediatype=1><img decoding=\"async\" class=\"lazyload html5galleryimg html5gallery-tn-image\" src=\"data:image\/svg+xml,%3Csvg%20xmlns%3D%27http%3A%2F%2Fwww.w3.org%2F2000%2Fsvg%27%20width%3D%271024%27%20height%3D%271024%27%20viewBox%3D%270%200%201024%201024%27%3E%3Crect%20width%3D%271024%27%20height%3D%271024%27%20fill-opacity%3D%220%22%2F%3E%3C%2Fsvg%3E\" data-orig-src=\"https:\/\/research.cchmc.org\/lungimage\/wp-content\/uploads\/2015\/04\/2014-004-014_C57Bl6_E16.5_LMM.14.24.4.53_SCGA1A1_CALCA_ACTA2-04.jpg\" alt=\"E16.5 Immunofluorescence and Confocal Imaging Showing Expression of CALCA, SCGB1A1, and ACTA2\" data-description=\"60X\"><\/a><div class=\"html5gallery-info\"><div class=\"html5gallery-title\">E16.5 Immunofluorescence and Confocal Imaging Showing Expression of CALCA, SCGB1A1, and ACTA2<\/div><div class=\"html5gallery-description\">60X<\/div><\/div><a class=\"html5galleryimglink\" href=\"https:\/\/research.cchmc.org\/lungimage\/wp-content\/uploads\/2015\/04\/2014-004-014_C57Bl6_E16.5_LMM.14.24.4.53_SCGA1A1_CALCA_ACTA2-05.jpg\" data-mediatype=1><img decoding=\"async\" class=\"lazyload html5galleryimg html5gallery-tn-image\" src=\"data:image\/svg+xml,%3Csvg%20xmlns%3D%27http%3A%2F%2Fwww.w3.org%2F2000%2Fsvg%27%20width%3D%271024%27%20height%3D%271024%27%20viewBox%3D%270%200%201024%201024%27%3E%3Crect%20width%3D%271024%27%20height%3D%271024%27%20fill-opacity%3D%220%22%2F%3E%3C%2Fsvg%3E\" data-orig-src=\"https:\/\/research.cchmc.org\/lungimage\/wp-content\/uploads\/2015\/04\/2014-004-014_C57Bl6_E16.5_LMM.14.24.4.53_SCGA1A1_CALCA_ACTA2-05.jpg\" alt=\"E16.5 Immunofluorescence and Confocal Imaging Showing Expression of CALCA, SCGB1A1, and ACTA2\" data-description=\"20X\"><\/a><div class=\"html5gallery-info\"><div class=\"html5gallery-title\">E16.5 Immunofluorescence and Confocal Imaging Showing Expression of CALCA, SCGB1A1, and ACTA2<\/div><div class=\"html5gallery-description\">20X<\/div><\/div><a class=\"html5galleryimglink\" href=\"https:\/\/research.cchmc.org\/lungimage\/wp-content\/uploads\/2015\/04\/2014-004-014_C57Bl6_E16.5_LMM.14.24.4.53_SCGA1A1_CALCA_ACTA2-06.jpg\" data-mediatype=1><img decoding=\"async\" class=\"lazyload html5galleryimg html5gallery-tn-image\" src=\"data:image\/svg+xml,%3Csvg%20xmlns%3D%27http%3A%2F%2Fwww.w3.org%2F2000%2Fsvg%27%20width%3D%271024%27%20height%3D%271024%27%20viewBox%3D%270%200%201024%201024%27%3E%3Crect%20width%3D%271024%27%20height%3D%271024%27%20fill-opacity%3D%220%22%2F%3E%3C%2Fsvg%3E\" data-orig-src=\"https:\/\/research.cchmc.org\/lungimage\/wp-content\/uploads\/2015\/04\/2014-004-014_C57Bl6_E16.5_LMM.14.24.4.53_SCGA1A1_CALCA_ACTA2-06.jpg\" alt=\"E16.5 Immunofluorescence and Confocal Imaging Showing Expression of CALCA, SCGB1A1, and ACTA2\" data-description=\"60X\"><\/a><div class=\"html5gallery-info\"><div class=\"html5gallery-title\">E16.5 Immunofluorescence and Confocal Imaging Showing Expression of CALCA, SCGB1A1, and ACTA2<\/div><div class=\"html5gallery-description\">60X<\/div><\/div><\/div><\/div><\/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<section class=\"elementor-section elementor-inner-section elementor-element elementor-element-5a68feb0 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"5a68feb0\" data-element_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-inner-column elementor-element elementor-element-5d731f30\" data-id=\"5d731f30\" data-element_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-234d83be elementor-widget elementor-widget-template\" data-id=\"234d83be\" data-element_type=\"widget\" data-widget_type=\"template.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<div class=\"elementor-template\">\n\t\t\t\t\t<div data-elementor-type=\"page\" data-elementor-id=\"24565\" class=\"elementor elementor-24565 elementor-bc-flex-widget\" data-elementor-post-type=\"elementor_library\">\n\t\t\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-4650a05 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"4650a05\" data-element_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-33 elementor-top-column elementor-element elementor-element-951322a\" data-id=\"951322a\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap\">\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t<div class=\"elementor-column elementor-col-33 elementor-top-column elementor-element elementor-element-4f951ba\" data-id=\"4f951ba\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap\">\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t<div class=\"elementor-column elementor-col-33 elementor-top-column elementor-element elementor-element-0022067\" data-id=\"0022067\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap\">\n\t\t\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\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<section class=\"elementor-section elementor-inner-section elementor-element elementor-element-ab47dc3 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"ab47dc3\" data-element_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-inner-column elementor-element elementor-element-4ee8cd7\" data-id=\"4ee8cd7\" data-element_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-5b5c7d89 elementor-widget elementor-widget-text-editor\" data-id=\"5b5c7d89\" data-element_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><strong>\u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0<\/strong><\/p><p><strong style=\"letter-spacing: 0px;\">Day 1<\/strong><\/p><ol><li>For Frozen tissue, rinse 2X in PBS, then 5 min in 4% PFA\/PBS, then rinse 1X in PBS.<\/li><li>Briefly equilibrate slides in Antigen Retrieval Buffer.<\/li><li>Antigen retrieval, pH 6.0 (times will vary according to microwave).<\/li><li>10 mM sodium citrate, pH 6.0, and heat in a microwave at 96<sup>o<\/sup>C.\u00a0*We usually do a series of three runs (6-7 minutes each run) to equal the time\/temp because of evaporation (refill coplin jars with dH<sub>2<\/sub>O).<\/li><li>Microwave according to instructions on microwave.<\/li><li>Cool on countertop, 15 min.<\/li><li>Rinse with dH<sub>2<\/sub>O<\/li><li>1X PBS, 5 min.<\/li><li>Block in 4% Goat serum\/PBS-T, 2 hours at RT.<\/li><li>For mouse anti \u2013<a href=\"http:\/\/www.sigmaaldrich.com\/catalog\/product\/sigma\/c7113?lang=en\u00aeion=US\" target=\"_Blank\"><strong>CALCA<\/strong><\/a> (CGRP) (C7113<strong>, Sigma<\/strong>)\u00a0dilute 1:200 in blocking buffer, For guinea pig anti-SCGB1A1 (G210, Bl. 3\/24\/11) dilute 1:1500, for mouse anti-\u00a0<a href=\"http:\/\/www.sigmaaldrich.com\/catalog\/product\/sigma\/a5228?lang=en\u00aeion=US\" target=\"_Blank\"><strong>ACTA2<\/strong><\/a>\u00a0 (\u03b1 Smooth muscle actin) (<a href=\"http:\/\/www.sigmaaldrich.com\/catalog\/product\/sigma\/a5228?lang=en\u00aeion=US\" target=\"_Blank\"><strong>A5228<\/strong><\/a>, Sigma)\u00a0dilute 1:2000 in blocking buffer. Spin for 10 minutes in a \u00b5fuge, apply to tissue, and incubate on tissue overnight @ 4<sup>o<\/sup>C.<\/li><\/ol><p><strong>Day 2<\/strong><\/p><ol><li>Rinse slides in PBS-T 3X, 5 min.<\/li><li>Apply secondary antibody , Goat Alexa Fluor 488 anti-mouse IgG1 (<strong><a href=\"http:\/\/www.lifetechnologies.com\/order\/catalog\/product\/A21121?ICID=search-a21121\" target=\"_Blank\">A21121<\/a><\/strong>, Lot# 1345050, for anti \u2013<a href=\"http:\/\/www.sigmaaldrich.com\/catalog\/product\/sigma\/c7113?lang=en\u00aeion=US\" target=\"_Blank\"><strong>CALCA<\/strong><\/a> (CGRP) ) dilute 1:200, Goat Alexa Fluor 555 anti-guinea pig IgG (<a href=\"http:\/\/www.lifetechnologies.com\/order\/catalog\/product\/A21435?ICID=search-a21435\" target=\"_Blank\"><strong>A21435<\/strong><\/a>, Lot# 1571699, anti-SCGBA1A1) at 1:200, Donkey Alexa Fluor 647 anti-mouse IgG2A (<strong><a href=\"https:\/\/www.lifetechnologies.com\/order\/catalog\/product\/A21136?CID=search-a21136\" target=\"_Blank\">A21136<\/a><\/strong>, Lot# 1345055, <a href=\"http:\/\/www.sigmaaldrich.com\/catalog\/product\/sigma\/a5228?lang=en\u00aeion=US\" target=\"_Blank\"><strong>ACTA2<\/strong><\/a>) at 1:200, in blocking buffer. Vortex and \u00b5fuge at full speed for 10 minutes. Incubated at room temperature for 1 hour.<\/li><li>Rinse in PBS-T 3X, 5 min.<\/li><li>Dilute DAPI 1:2000 and apply to slides for 10 min.<\/li><li>Wash in PBS-T 3X, 5 min.<\/li><li>Rinse slides in 0.1M PB or TB, 3X, 5 min.<\/li><li>Add 1 drop of Prolong Gold anti-fade mounting medium (P36930).<\/li><li>Coverslip with Gold Seal Coverslip (Cat# 3422 Electron Microscopy Sciences, 22 X 22 mm).<\/li><li>Allow Prolong Gold to cure overnight at room temp in light sealed box.<\/li><li>Store slides in a light sealed box @ 4<sup>o<\/sup>C.<\/li><\/ol><p><strong>\u00a0<\/strong><\/p><p><strong>Tissue Used: \u00a0\u00a0<\/strong><\/p><p>LMM.14.24.4.53 E16.5<\/p><p>Gender: Male<\/p><p>Crown-to-Rump Length: 16.0 mm<\/p><p>Weight=0.592g<\/p><p><strong>\u00a0<\/strong><\/p><p><strong>Appendix<\/strong><\/p><p><strong>Antigen Retrieval Solution<\/strong><\/p><p>18ml Solution A- 0.1M Citric Acid (Sigma, C1909) pH 2.5<\/p><p>82ml Reagent B- 0.1M Sodium Citrate (Sigma, S4641 ) pH 8.2<\/p><p>1L dH<sub>2<\/sub>O<\/p><p>pH 6.0<\/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-inner-section elementor-element elementor-element-5928b96a elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"5928b96a\" data-element_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-inner-column elementor-element elementor-element-60ce0b23\" data-id=\"60ce0b23\" data-element_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-2c0a8b8f elementor-widget elementor-widget-facebook-comments\" data-id=\"2c0a8b8f\" data-element_type=\"widget\" data-widget_type=\"facebook-comments.default\">\n\t\t\t\t<div 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id=\"recent-posts-33\" class=\"widget widget_recent_entries\">\n\t\t<div class=\"heading\"><h4 class=\"widget-title\">Recent Experiments<\/h4><\/div>\n\t\t<ul>\n\t\t\t\t\t\t\t\t\t\t\t<li>\n\t\t\t\t\t<a href=\"https:\/\/research.cchmc.org\/lungimage\/?p=32066\">4 Year-Old Human Lung KRT14, MUC5B, and TUBA4A Confocal Imaging<\/a>\n\t\t\t\t\t\t\t\t\t<\/li>\n\t\t\t\t\t\t\t\t\t\t\t<li>\n\t\t\t\t\t<a href=\"https:\/\/research.cchmc.org\/lungimage\/?p=31223\">Alcian Blue Staining from PBR 3653 Patient with Cystic Fibrosis<\/a>\n\t\t\t\t\t\t\t\t\t<\/li>\n\t\t\t\t\t\t\t\t\t\t\t<li>\n\t\t\t\t\t<a href=\"https:\/\/research.cchmc.org\/lungimage\/?p=31202\">H&#038;E Staining from PBR 3383 Patient with Cystic Fibrosis<\/a>\n\t\t\t\t\t\t\t\t\t<\/li>\n\t\t\t\t\t\t\t\t\t\t\t<li>\n\t\t\t\t\t<a href=\"https:\/\/research.cchmc.org\/lungimage\/?p=31199\">H&#038;E Staining from PBR 3653 Patient with Cystic Fibrosis<\/a>\n\t\t\t\t\t\t\t\t\t<\/li>\n\t\t\t\t\t\t\t\t\t\t\t<li>\n\t\t\t\t\t<a href=\"https:\/\/research.cchmc.org\/lungimage\/?p=31196\">H&#038;E Staining of 31 Year-Old Human Donor 10<\/a>\n\t\t\t\t\t\t\t\t\t<\/li>\n\t\t\t\t\t\t\t\t\t\t\t<li>\n\t\t\t\t\t<a href=\"https:\/\/research.cchmc.org\/lungimage\/?p=31193\">H&#038;E Staining of 27 Year-Old Human Donor 3<\/a>\n\t\t\t\t\t\t\t\t\t<\/li>\n\t\t\t\t\t\t\t\t\t\t\t<li>\n\t\t\t\t\t<a href=\"https:\/\/research.cchmc.org\/lungimage\/?p=31190\">Alcian Blue Staining of 27 Year-Old Human Donor 3<\/a>\n\t\t\t\t\t\t\t\t\t<\/li>\n\t\t\t\t\t\t\t\t\t\t\t<li>\n\t\t\t\t\t<a href=\"https:\/\/research.cchmc.org\/lungimage\/?p=31186\">Alcian Blue Staining of 31 Year-Old Human Donor 10<\/a>\n\t\t\t\t\t\t\t\t\t<\/li>\n\t\t\t\t\t\t\t\t\t\t\t<li>\n\t\t\t\t\t<a href=\"https:\/\/research.cchmc.org\/lungimage\/?p=31182\">Alcian Blue Staining from PBR 3383 Patient with Cystic Fibrosis<\/a>\n\t\t\t\t\t\t\t\t\t<\/li>\n\t\t\t\t\t\t\t\t\t\t\t<li>\n\t\t\t\t\t<a href=\"https:\/\/research.cchmc.org\/lungimage\/?p=31024\">Alcian Blue Staining of 29 Year-Old Human Donor D175-10C1.21<\/a>\n\t\t\t\t\t\t\t\t\t<\/li>\n\t\t\t\t\t\t\t\t\t\t\t<li>\n\t\t\t\t\t<a href=\"https:\/\/research.cchmc.org\/lungimage\/?p=31019\">Alcian Blue Staining of 29 Year-Old Human Donor D175-11C4.14<\/a>\n\t\t\t\t\t\t\t\t\t<\/li>\n\t\t\t\t\t\t\t\t\t\t\t<li>\n\t\t\t\t\t<a href=\"https:\/\/research.cchmc.org\/lungimage\/?p=31016\">Alcian Blue Staining of 29 Year-Old Human Donor D175-11C2.11<\/a>\n\t\t\t\t\t\t\t\t\t<\/li>\n\t\t\t\t\t\t\t\t\t\t\t<li>\n\t\t\t\t\t<a href=\"https:\/\/research.cchmc.org\/lungimage\/?p=31013\">Alcian Blue Staining of 24 Year-Old Human Donor D071-23C2.19<\/a>\n\t\t\t\t\t\t\t\t\t<\/li>\n\t\t\t\t\t\t\t\t\t\t\t<li>\n\t\t\t\t\t<a href=\"https:\/\/research.cchmc.org\/lungimage\/?p=31010\">Alcian Blue Staining of 24 Year-Old Human Donor D071-17C2.20<\/a>\n\t\t\t\t\t\t\t\t\t<\/li>\n\t\t\t\t\t\t\t\t\t\t\t<li>\n\t\t\t\t\t<a href=\"https:\/\/research.cchmc.org\/lungimage\/?p=31007\">Alcian Blue Staining of 24 Year-Old Human Donor D071-16C3.27<\/a>\n\t\t\t\t\t\t\t\t\t<\/li>\n\t\t\t\t\t<\/ul>\n\n\t\t<\/div>\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>\u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 Day 1 For Frozen tissue, rinse 2X in PBS, then 5 min in 4% PFA\/PBS, then rinse 1X in PBS. Briefly equilibrate slides [&hellip;]<\/p>\n","protected":false},"author":1,"featured_media":2622,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[81,274],"tags":[],"class_list":["post-634","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-e16-5","category-c57bl6"],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v25.6 - https:\/\/yoast.com\/wordpress\/plugins\/seo\/ -->\n<title>E16.5 C57BL6 SCGB1A1, CALCA, and ACTA2 Confocal Imaging -<\/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:\/\/research.cchmc.org\/lungimage\/?p=634\" \/>\n<meta property=\"og:locale\" content=\"en_US\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"E16.5 C57BL6 SCGB1A1, CALCA, and ACTA2 Confocal Imaging -\" \/>\n<meta property=\"og:description\" content=\"\u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 Day 1 For Frozen tissue, rinse 2X in PBS, then 5 min in 4% PFA\/PBS, then rinse 1X in PBS. 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