{"id":19234,"date":"2018-02-20T20:14:41","date_gmt":"2018-02-20T20:14:41","guid":{"rendered":"https:\/\/research.cchmc.org\/lungimage\/?p=19234"},"modified":"2018-02-26T17:09:49","modified_gmt":"2018-02-26T17:09:49","slug":"plish-in-situ-hybridization-of-200-%c2%b5m-human-lung-tissue-cleared-in-scale-for-sftpc-mir29-1-and-axin2","status":"publish","type":"post","link":"https:\/\/research.cchmc.org\/lungimage\/?p=19234","title":{"rendered":"PLISH In situ Hybridization of 200 \u00b5m Human Lung Tissue PACT for SFTPC"},"content":{"rendered":"<div class=\"fusion-fullwidth fullwidth-box fusion-builder-row-1 hundred-percent-fullwidth non-hundred-percent-height-scrolling\" style=\"--awb-border-radius-top-left:0px;--awb-border-radius-top-right:0px;--awb-border-radius-bottom-right:0px;--awb-border-radius-bottom-left:0px;--awb-overflow:visible;--awb-flex-wrap:wrap;\" ><div class=\"fusion-builder-row fusion-row\"><div class=\"fusion-layout-column fusion_builder_column fusion-builder-column-0 fusion_builder_column_1_1 1_1 fusion-one-full fusion-column-first fusion-column-last\" style=\"--awb-bg-size:cover;--awb-margin-bottom:25px;\"><div class=\"fusion-column-wrapper fusion-flex-column-wrapper-legacy\"><div class=\"fusion-text fusion-text-1\"><\/div><div class=\"fusion-text fusion-text-2\"><div class=\"wonderplugingallery-container\" id=\"wonderplugingallery-container-670\" style=\"max-width:550px;margin:0 auto;\"><div class=\"wonderplugingallery\" id=\"wonderplugingallery-670\" data-galleryid=\"670\" data-width=\"550\" data-height=\"550\" data-skin=\"light\" data-random=\"false\" data-autoslide=\"false\" data-autoplayvideo=\"false\" data-schemamarkup=\"false\" data-stopallplaying=\"false\" data-reloadonvideoend=\"false\" data-enabletabindex=\"false\" data-loadnextonvideoend=\"false\" data-hidetitlewhenvideoisplaying=\"false\" data-disablehovereventontouch=\"false\" data-autoslideandplayafterfirstplayed=\"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-donotinit=\"false\" data-addinitscript=\"false\" data-triggerresize=\"false\" data-thumbcolumnsresponsive=\"false\" data-showimgtitle=\"false\" data-titlesmallscreen=\"false\" data-initsocial=\"true\" data-showsocial=\"false\" data-showemail=\"false\" data-showfacebook=\"true\" data-showtwitter=\"true\" data-showpinterest=\"true\" data-socialrotateeffect=\"true\" data-doshortcodeontext=\"false\" data-duration=\"1500\" data-slideduration=\"1000\" data-slideshowinterval=\"6000\" 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-titleheight=\"72\" data-titlesmallscreenwidth=\"640\" data-titleheightsmallscreen=\"148\" data-socialmode=\"mouseover\" data-socialposition=\"position:absolute;top:8px;right:8px;\" data-socialpositionlightbox=\"position:absolute;top:8px;right:8px;\" data-socialdirection=\"horizontal\" data-socialbuttonsize=\"32\" data-socialbuttonfontsize=\"18\" data-triggerresizedelay=\"100\" data-thumbmediumsize=\"800\" data-thumbsmallsize=\"480\" data-thumbmediumwidth=\"64\" data-thumbmediumheight=\"64\" data-thumbsmallwidth=\"48\" data-thumbsmallheight=\"48\" data-imgtitle=\"title\" data-jsfolder=\"https:\/\/research.cchmc.org\/lungimage\/wp-content\/plugins\/wonderplugin-gallery\/engine\/\" style=\"display:none;\" ><a class=\"html5galleryimglink\" href=\"https:\/\/www.youtube.com\/embed\/9kReDWsvVV8?t=10s\" data-poster=\"https:\/\/img.youtube.com\/vi\/9kReDWsvVV8\/0.jpg\"><img decoding=\"async\" class=\"lazyload html5galleryimg html5gallery-tn-image\" src=\"data:image\/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==\" data-orig-src=\"https:\/\/img.youtube.com\/vi\/9kReDWsvVV8\/0.jpg\" alt=\"PLISH In situ hybridization of thick human lung tissue For SFTPC, MIR29-1, and AXIN2\" data-description=\"Donor DD036\n40X\"><\/a><div class=\"html5gallery-info\"><div class=\"html5gallery-title\">PLISH In situ hybridization of thick human lung tissue For SFTPC, MIR29-1, and AXIN2<\/div><div class=\"html5gallery-description\">Donor DD036\n40X<\/div><\/div><a class=\"html5galleryimglink\" href=\"https:\/\/www.youtube.com\/embed\/6O_kfcAg4nk\" data-poster=\"https:\/\/img.youtube.com\/vi\/6O_kfcAg4nk\/0.jpg\"><img decoding=\"async\" class=\"lazyload html5galleryimg html5gallery-tn-image\" src=\"data:image\/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==\" data-orig-src=\"https:\/\/img.youtube.com\/vi\/6O_kfcAg4nk\/0.jpg\" alt=\"PLISH In situ hybridization of thick human lung tissue For SFTPC, MIR29-1, and AXIN2\" data-description=\"Donor DD036\n40X\"><\/a><div class=\"html5gallery-info\"><div class=\"html5gallery-title\">PLISH In situ hybridization of thick human lung tissue For SFTPC, MIR29-1, and AXIN2<\/div><div class=\"html5gallery-description\">Donor DD036\n40X<\/div><\/div><a class=\"html5galleryimglink\" href=\"https:\/\/www.youtube.com\/embed\/fmHaAsH3xGw\" data-poster=\"https:\/\/img.youtube.com\/vi\/fmHaAsH3xGw\/0.jpg\"><img decoding=\"async\" class=\"lazyload html5galleryimg html5gallery-tn-image\" src=\"data:image\/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==\" data-orig-src=\"https:\/\/img.youtube.com\/vi\/fmHaAsH3xGw\/0.jpg\" alt=\"PLISH In situ hybridization of thick human lung tissue For SFTPC, MIR29-1, and AXIN2\" data-description=\"Donor DD036\n40X\"><\/a><div class=\"html5gallery-info\"><div class=\"html5gallery-title\">PLISH In situ hybridization of thick human lung tissue For SFTPC, MIR29-1, and AXIN2<\/div><div class=\"html5gallery-description\">Donor DD036\n40X<\/div><\/div><a class=\"html5galleryimglink\" href=\"https:\/\/www.youtube.com\/embed\/xjN-XJjjutw\" data-poster=\"https:\/\/img.youtube.com\/vi\/xjN-XJjjutw\/0.jpg\"><img decoding=\"async\" class=\"lazyload html5galleryimg html5gallery-tn-image\" src=\"data:image\/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==\" data-orig-src=\"https:\/\/img.youtube.com\/vi\/xjN-XJjjutw\/0.jpg\" alt=\"PLISH In situ hybridization of thick human lung tissue For SFTPC, MIR29-1, and AXIN2\" data-description=\"Donor DD036\n40X\"><\/a><div class=\"html5gallery-info\"><div class=\"html5gallery-title\">PLISH In situ hybridization of thick human lung tissue For SFTPC, MIR29-1, and AXIN2<\/div><div class=\"html5gallery-description\">Donor DD036\n40X<\/div><\/div><a class=\"html5galleryimglink\" href=\"https:\/\/www.youtube.com\/embed\/m8dXjo_rVGg\" data-poster=\"https:\/\/img.youtube.com\/vi\/m8dXjo_rVGg\/0.jpg\"><img decoding=\"async\" class=\"lazyload html5galleryimg html5gallery-tn-image\" src=\"data:image\/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==\" data-orig-src=\"https:\/\/img.youtube.com\/vi\/m8dXjo_rVGg\/0.jpg\" alt=\"PLISH In situ hybridization of thick human lung tissue For SFTPC, MIR29-1, and AXIN2\" data-description=\"Donor DD036\n40X\"><\/a><div class=\"html5gallery-info\"><div class=\"html5gallery-title\">PLISH In situ hybridization of thick human lung tissue For SFTPC, MIR29-1, and AXIN2<\/div><div class=\"html5gallery-description\">Donor DD036\n40X<\/div><\/div><a class=\"html5galleryimglink\" href=\"https:\/\/www.youtube.com\/embed\/a_qrFaWQhi4\" data-poster=\"https:\/\/img.youtube.com\/vi\/a_qrFaWQhi4\/0.jpg\"><img decoding=\"async\" class=\"lazyload html5galleryimg html5gallery-tn-image\" src=\"data:image\/gif;base64,R0lGODlhAQABAAAAACH5BAEKAAEALAAAAAABAAEAAAICTAEAOw==\" data-orig-src=\"https:\/\/img.youtube.com\/vi\/a_qrFaWQhi4\/0.jpg\" alt=\"PLISH In situ hybridization of thick human lung tissue For SFTPC, MIR29-1, and AXIN2\" data-description=\"Donor DD036\n40X\"><\/a><div class=\"html5gallery-info\"><div class=\"html5gallery-title\">PLISH In situ hybridization of thick human lung tissue For SFTPC, MIR29-1, and AXIN2<\/div><div class=\"html5gallery-description\">Donor DD036\n40X<\/div><\/div><\/div><\/div>\n<\/div><div class=\"fusion-clearfix\"><\/div><\/div><\/div><div class=\"fusion-layout-column fusion_builder_column fusion-builder-column-1 fusion_builder_column_1_1 1_1 fusion-one-full fusion-column-first fusion-column-last\" style=\"--awb-bg-size:cover;--awb-margin-bottom:25px;\"><div class=\"fusion-column-wrapper fusion-flex-column-wrapper-legacy\"><div class=\"fusion-text fusion-text-3\"><\/div><div class=\"fusion-text fusion-text-4\"><table class=\" aligncenter\" style=\"height: 26px; background-color: #030000;\" width=\"262\">\n<tbody>\n<tr>\n<td style=\"width: 80.1136px; text-align: center;\"><strong><span style=\"color: #339966;\">SFTPC<\/span><\/strong><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>Donor tissue kindly provided by <strong>Dr. Scott Randell, <a href=\"https:\/\/www.med.unc.edu\/marsicolunginstitute\/directory\/scott-h-randell-phd\/\" target=\"_blank\" rel=\"noopener\">University of North Carolina<\/a><\/strong><\/p>\n<p><strong>Purpose:\u00a0 Plish in situ hybridization on PACT cleared tissue for SFTPC, MIR29-1, and AXIN2<\/strong><\/p>\n<\/div><div class=\"fusion-tabs fusion-tabs-1 classic vertical-tabs icon-position-left mobile-mode-accordion\" style=\"--awb-title-border-radius-top-left:0px;--awb-title-border-radius-top-right:0px;--awb-title-border-radius-bottom-right:0px;--awb-title-border-radius-bottom-left:0px;--awb-inactive-color:#ebeaea;--awb-background-color:#ffffff;--awb-border-color:#ebeaea;--awb-active-border-color:#1a80b6;\"><div class=\"nav\"><ul class=\"nav-tabs\" role=\"tablist\"><li class=\"active\" role=\"presentation\"><a class=\"tab-link\" data-toggle=\"tab\" role=\"tab\" aria-controls=\"tab-ea55814d540a26ee79e\" aria-selected=\"true\" id=\"fusion-tab-tissuepreparation\" href=\"#tab-ea55814d540a26ee79e\"><h4 class=\"fusion-tab-heading\">Tissue Preparation<\/h4><\/a><\/li><li role=\"presentation\"><a class=\"tab-link\" data-toggle=\"tab\" role=\"tab\" aria-controls=\"tab-3452deb7662bb36f8ac\" aria-selected=\"false\" tabindex=\"-1\" id=\"fusion-tab-pactclearing\" href=\"#tab-3452deb7662bb36f8ac\"><h4 class=\"fusion-tab-heading\">PACT Clearing<\/h4><\/a><\/li><li role=\"presentation\"><a class=\"tab-link\" data-toggle=\"tab\" role=\"tab\" aria-controls=\"tab-32099aea25199518b52\" aria-selected=\"false\" tabindex=\"-1\" id=\"fusion-tab-plish\" href=\"#tab-32099aea25199518b52\"><h4 class=\"fusion-tab-heading\">PLISH<\/h4><\/a><\/li><li role=\"presentation\"><a class=\"tab-link\" data-toggle=\"tab\" role=\"tab\" aria-controls=\"tab-303ecf8cce992e98406\" aria-selected=\"false\" tabindex=\"-1\" id=\"fusion-tab-references\" href=\"#tab-303ecf8cce992e98406\"><h4 class=\"fusion-tab-heading\">References<\/h4><\/a><\/li><li role=\"presentation\"><a class=\"tab-link\" data-toggle=\"tab\" role=\"tab\" aria-controls=\"tab-ee1d272236f0ec01101\" aria-selected=\"false\" tabindex=\"-1\" id=\"fusion-tab-tissuemetadata\" href=\"#tab-ee1d272236f0ec01101\"><h4 class=\"fusion-tab-heading\">Tissue Metadata<\/h4><\/a><\/li><\/ul><\/div><div class=\"tab-content\"><div class=\"nav fusion-mobile-tab-nav\"><ul class=\"nav-tabs\" role=\"tablist\"><li class=\"active\" role=\"presentation\"><a class=\"tab-link\" data-toggle=\"tab\" role=\"tab\" aria-controls=\"tab-ea55814d540a26ee79e\" aria-selected=\"true\" id=\"mobile-fusion-tab-tissuepreparation\" href=\"#tab-ea55814d540a26ee79e\"><h4 class=\"fusion-tab-heading\">Tissue Preparation<\/h4><\/a><\/li><\/ul><\/div><div class=\"tab-pane fade fusion-clearfix in active\" role=\"tabpanel\" tabindex=\"0\" aria-labelledby=\"fusion-tab-tissuepreparation\" id=\"tab-ea55814d540a26ee79e\">\n<p>Human lung tissue (kindly provided by Dr. Scott Randell) was fixed in 10% formalin. Tissue was then washed in DEPC-treated 1X PBS and incubated in 30% Sucrose in DEPC-treated 1X PBS. 24 hours prior to cryoprotection, tissue was placed in Sucrose\/OCT (2:1). Upon freezing, lung tissue was placed in OCT and frozen on dry ice and stored at -80<sup>o<\/sup>C. 200 \u00b5m sections of OCT protected frozen human lung were sectioned on a cryostat and stored at -20<sup>o<\/sup>C until clearing.<\/p>\n<\/div><div class=\"nav fusion-mobile-tab-nav\"><ul class=\"nav-tabs\" role=\"tablist\"><li role=\"presentation\"><a class=\"tab-link\" data-toggle=\"tab\" role=\"tab\" aria-controls=\"tab-3452deb7662bb36f8ac\" aria-selected=\"false\" tabindex=\"-1\" id=\"mobile-fusion-tab-pactclearing\" href=\"#tab-3452deb7662bb36f8ac\"><h4 class=\"fusion-tab-heading\">PACT Clearing<\/h4><\/a><\/li><\/ul><\/div><div class=\"tab-pane fade fusion-clearfix\" role=\"tabpanel\" tabindex=\"0\" aria-labelledby=\"fusion-tab-pactclearing\" id=\"tab-3452deb7662bb36f8ac\">\n<p>Thick sections were rehydrated in 1X PBS and washed several times to remove OCT. Lungs were incubated in 4% Acrylamide\/0.25% 2,2\u2019-Azobis[2-(2-imidazolin-2-yl)propane]dihydrochloride photoinitiator (VA-044, Wako) in 1X PBS overnight at 4<sup>o<\/sup>C in a 50ml conical tube. Thick lung sections were then polymerized for 3 hours at 37<sup>o<\/sup>C. Following polymerization, excess hydrogel was washed away with several washes of 1X PBS. PBS was removed, and the thick sections were incubated in 8% SDS in 1X PBS overnight in a shaking incubator at 37<sup>o<\/sup>C. Thick lung sections were washed several times in 1X PBS to completely remove SDS.<\/p>\n<\/div><div class=\"nav fusion-mobile-tab-nav\"><ul class=\"nav-tabs\" role=\"tablist\"><li role=\"presentation\"><a class=\"tab-link\" data-toggle=\"tab\" role=\"tab\" aria-controls=\"tab-32099aea25199518b52\" aria-selected=\"false\" tabindex=\"-1\" id=\"mobile-fusion-tab-plish\" href=\"#tab-32099aea25199518b52\"><h4 class=\"fusion-tab-heading\">PLISH<\/h4><\/a><\/li><\/ul><\/div><div class=\"tab-pane fade fusion-clearfix\" role=\"tabpanel\" tabindex=\"0\" aria-labelledby=\"fusion-tab-plish\" id=\"tab-32099aea25199518b52\">\n<div class=\"_rp_l\" role=\"heading\" aria-level=\"2\">\n<div class=\"ms-font-color-neutralPrimary ms-font-xl ms-fwt-r _rp_m\">\n<p>Hybridization probes were incubated at concentrations of 100 nM each in hybridization buffer (1M NaTCA, 5mM EDTA, 50mM Tris pH7.4, 0.2mg.mL Heparin in DEPC water) for two hours at 37\u00baC and 100% humidity. The sections were then incubated with ligase buffer and phosphorylated common bridge and connector circle oligos at 10 nM for 60 minutes and then with ligase buffer, T4 ligase, and ligase buffer for two hours under the same conditions. DNA amplification was then accomplished using Phi-29 polymerase (Lucigen-30221) in polymerase buffer at the same conditions overnight. After the reaction was complete, slides were washed with label buffer (2X SCC\/20% Formamide in DEPC water) and then incubated with 100nM Tye705 label probe in label probe buffer for one hour in the same conditions. Immunofluorescent co-staining and imaging were obtained and analyzed as follows. Negative controls of secondary antibody alone and reactions performed with identically labeled probes for the bacteria Bacillus subtilis gene mgsA were imaged and used for thresholding for quantitative image analysis. All probes were ordered from Integrated DNA Technologies, target probes were ordered with standard desalting and all probes were stored as 100\u03bcM stocks.<\/p>\n<\/div>\n<\/div>\n<\/div><div class=\"nav fusion-mobile-tab-nav\"><ul class=\"nav-tabs\" role=\"tablist\"><li role=\"presentation\"><a class=\"tab-link\" data-toggle=\"tab\" role=\"tab\" aria-controls=\"tab-303ecf8cce992e98406\" aria-selected=\"false\" tabindex=\"-1\" id=\"mobile-fusion-tab-references\" href=\"#tab-303ecf8cce992e98406\"><h4 class=\"fusion-tab-heading\">References<\/h4><\/a><\/li><\/ul><\/div><div class=\"tab-pane fade fusion-clearfix\" role=\"tabpanel\" tabindex=\"0\" aria-labelledby=\"fusion-tab-references\" id=\"tab-303ecf8cce992e98406\">\n<p><strong>Single-Cell Phenotyping within Transparent Intact Tissue through Whole-Body Clearing<\/strong><br \/>\nYang, Bin et al. Cell , Volume 158 , Issue 4 , 945 &#8211; 958<br \/>\n<u><a href=\"http:\/\/dx.doi.org\/10.1016\/j.cell.2014.07.017\">http:\/\/dx.doi.org\/10.1016\/j.cell.2014.07.017<\/a><\/u><\/p>\n<p><strong>Automated cell-type classification in intact tissues by single-cell molecular profiling<\/strong><br \/>\n<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/29319504\">Elife.<\/a>\u00a02018 Jan 10;7. pii: e30510. doi: 10.7554\/eLife.30510<br \/>\n<u><a href=\"https:\/\/elifesciences.org\/articles\/30510\">https:\/\/elifesciences.org\/articles\/30510<\/a><\/u><\/p>\n<p><strong>Single-cell Wnt signaling niches maintain stemness of alveolar type 2 cells<\/strong><br \/>\n<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pubmed\/?term=Single-cell+Wnt+signaling+niches+maintain+stemness+of+alveolar+type+2+cells\">Science.<\/a>\u00a02018 Feb 1. pii: eaam6603. doi: 10.1126\/science.aam6603<br \/>\n<a href=\"http:\/\/science.sciencemag.org\/content\/early\/2018\/01\/31\/science.aam6603.long\" target=\"_blank\" rel=\"noopener\"><u>http:\/\/science.sciencemag.org\/content\/early\/2018\/01\/31\/science.aam6603.long<\/u><\/a><\/p>\n<\/div><div class=\"nav fusion-mobile-tab-nav\"><ul class=\"nav-tabs\" role=\"tablist\"><li role=\"presentation\"><a class=\"tab-link\" data-toggle=\"tab\" role=\"tab\" aria-controls=\"tab-ee1d272236f0ec01101\" aria-selected=\"false\" tabindex=\"-1\" id=\"mobile-fusion-tab-tissuemetadata\" href=\"#tab-ee1d272236f0ec01101\"><h4 class=\"fusion-tab-heading\">Tissue Metadata<\/h4><\/a><\/li><\/ul><\/div><div class=\"tab-pane fade fusion-clearfix\" role=\"tabpanel\" tabindex=\"0\" aria-labelledby=\"fusion-tab-tissuemetadata\" id=\"tab-ee1d272236f0ec01101\">\n<p><strong>LMHA-15-UNC-DD036L_MC15-04H.4.1<\/strong><\/p>\n<p><strong>Gender: Male<\/strong><\/p>\n<p><strong>Age:\u00a031 Year Old<\/strong><\/p>\n<p><strong>Race: Hispanic<\/strong><\/p>\n<p><strong>Non-Smoker<\/strong><\/p>\n<\/div><\/div><\/div><div class=\"fusion-clearfix\"><\/div><\/div><\/div><\/div><\/div>\n","protected":false},"excerpt":{"rendered":"","protected":false},"author":1,"featured_media":0,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[1],"tags":[],"class_list":["post-19234","post","type-post","status-publish","format-standard","hentry","category-uncategorized"],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v25.6 - https:\/\/yoast.com\/wordpress\/plugins\/seo\/ -->\n<title>PLISH In situ Hybridization of 200 \u00b5m Human Lung Tissue PACT for SFTPC -<\/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=19234\" \/>\n<meta property=\"og:locale\" content=\"en_US\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"PLISH In situ Hybridization of 200 \u00b5m Human Lung Tissue PACT for SFTPC -\" \/>\n<meta property=\"og:url\" content=\"https:\/\/research.cchmc.org\/lungimage\/?p=19234\" \/>\n<meta property=\"article:published_time\" content=\"2018-02-20T20:14:41+00:00\" \/>\n<meta property=\"article:modified_time\" content=\"2018-02-26T17:09:49+00:00\" \/>\n<meta name=\"author\" content=\"kitc5i\" \/>\n<meta name=\"twitter:card\" content=\"summary_large_image\" \/>\n<meta name=\"twitter:label1\" content=\"Written by\" \/>\n\t<meta name=\"twitter:data1\" content=\"kitc5i\" \/>\n\t<meta name=\"twitter:label2\" content=\"Est. reading time\" \/>\n\t<meta name=\"twitter:data2\" content=\"4 minutes\" \/>\n<script type=\"application\/ld+json\" class=\"yoast-schema-graph\">{\"@context\":\"https:\/\/schema.org\",\"@graph\":[{\"@type\":\"Article\",\"@id\":\"https:\/\/research.cchmc.org\/lungimage\/?p=19234#article\",\"isPartOf\":{\"@id\":\"https:\/\/research.cchmc.org\/lungimage\/?p=19234\"},\"author\":{\"name\":\"kitc5i\",\"@id\":\"https:\/\/research.cchmc.org\/lungimage\/#\/schema\/person\/94220f9638d78ff90fe4cd1d5743081f\"},\"headline\":\"PLISH In situ Hybridization of 200 \u00b5m Human Lung Tissue PACT for SFTPC\",\"datePublished\":\"2018-02-20T20:14:41+00:00\",\"dateModified\":\"2018-02-26T17:09:49+00:00\",\"mainEntityOfPage\":{\"@id\":\"https:\/\/research.cchmc.org\/lungimage\/?p=19234\"},\"wordCount\":724,\"publisher\":{\"@id\":\"https:\/\/research.cchmc.org\/lungimage\/#organization\"},\"inLanguage\":\"en-US\"},{\"@type\":\"WebPage\",\"@id\":\"https:\/\/research.cchmc.org\/lungimage\/?p=19234\",\"url\":\"https:\/\/research.cchmc.org\/lungimage\/?p=19234\",\"name\":\"PLISH In situ Hybridization of 200 \u00b5m Human Lung Tissue PACT for SFTPC -\",\"isPartOf\":{\"@id\":\"https:\/\/research.cchmc.org\/lungimage\/#website\"},\"datePublished\":\"2018-02-20T20:14:41+00:00\",\"dateModified\":\"2018-02-26T17:09:49+00:00\",\"breadcrumb\":{\"@id\":\"https:\/\/research.cchmc.org\/lungimage\/?p=19234#breadcrumb\"},\"inLanguage\":\"en-US\",\"potentialAction\":[{\"@type\":\"ReadAction\",\"target\":[\"https:\/\/research.cchmc.org\/lungimage\/?p=19234\"]}]},{\"@type\":\"BreadcrumbList\",\"@id\":\"https:\/\/research.cchmc.org\/lungimage\/?p=19234#breadcrumb\",\"itemListElement\":[{\"@type\":\"ListItem\",\"position\":1,\"name\":\"Home\",\"item\":\"https:\/\/research.cchmc.org\/lungimage\/\"},{\"@type\":\"ListItem\",\"position\":2,\"name\":\"PLISH In situ Hybridization of 200 \u00b5m Human Lung Tissue PACT for SFTPC\"}]},{\"@type\":\"WebSite\",\"@id\":\"https:\/\/research.cchmc.org\/lungimage\/#website\",\"url\":\"https:\/\/research.cchmc.org\/lungimage\/\",\"name\":\"\",\"description\":\"Lung Images\",\"publisher\":{\"@id\":\"https:\/\/research.cchmc.org\/lungimage\/#organization\"},\"potentialAction\":[{\"@type\":\"SearchAction\",\"target\":{\"@type\":\"EntryPoint\",\"urlTemplate\":\"https:\/\/research.cchmc.org\/lungimage\/?s={search_term_string}\"},\"query-input\":{\"@type\":\"PropertyValueSpecification\",\"valueRequired\":true,\"valueName\":\"search_term_string\"}}],\"inLanguage\":\"en-US\"},{\"@type\":\"Organization\",\"@id\":\"https:\/\/research.cchmc.org\/lungimage\/#organization\",\"name\":\"Cincinnati Children's Hospital Medical Center\",\"url\":\"https:\/\/research.cchmc.org\/lungimage\/\",\"logo\":{\"@type\":\"ImageObject\",\"inLanguage\":\"en-US\",\"@id\":\"https:\/\/research.cchmc.org\/lungimage\/#\/schema\/logo\/image\/\",\"url\":\"https:\/\/research.cchmc.org\/lungimage\/wp-content\/uploads\/2018\/05\/CCHMC-LOGO-II.png\",\"contentUrl\":\"https:\/\/research.cchmc.org\/lungimage\/wp-content\/uploads\/2018\/05\/CCHMC-LOGO-II.png\",\"width\":668,\"height\":277,\"caption\":\"Cincinnati Children's Hospital Medical Center\"},\"image\":{\"@id\":\"https:\/\/research.cchmc.org\/lungimage\/#\/schema\/logo\/image\/\"}},{\"@type\":\"Person\",\"@id\":\"https:\/\/research.cchmc.org\/lungimage\/#\/schema\/person\/94220f9638d78ff90fe4cd1d5743081f\",\"name\":\"kitc5i\",\"image\":{\"@type\":\"ImageObject\",\"inLanguage\":\"en-US\",\"@id\":\"https:\/\/research.cchmc.org\/lungimage\/#\/schema\/person\/image\/\",\"url\":\"https:\/\/secure.gravatar.com\/avatar\/e4bb6585a3f4dcb372eaf2f60ace1104aa6c71adfb2a1fa31533d31bb61f4034?s=96&d=mm&r=g\",\"contentUrl\":\"https:\/\/secure.gravatar.com\/avatar\/e4bb6585a3f4dcb372eaf2f60ace1104aa6c71adfb2a1fa31533d31bb61f4034?s=96&d=mm&r=g\",\"caption\":\"kitc5i\"},\"url\":\"https:\/\/research.cchmc.org\/lungimage\/?author=1\"}]}<\/script>\n<!-- \/ Yoast SEO plugin. -->","yoast_head_json":{"title":"PLISH In situ Hybridization of 200 \u00b5m Human Lung Tissue PACT for SFTPC -","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:\/\/research.cchmc.org\/lungimage\/?p=19234","og_locale":"en_US","og_type":"article","og_title":"PLISH In situ Hybridization of 200 \u00b5m Human Lung Tissue PACT for SFTPC -","og_url":"https:\/\/research.cchmc.org\/lungimage\/?p=19234","article_published_time":"2018-02-20T20:14:41+00:00","article_modified_time":"2018-02-26T17:09:49+00:00","author":"kitc5i","twitter_card":"summary_large_image","twitter_misc":{"Written by":"kitc5i","Est. reading time":"4 minutes"},"schema":{"@context":"https:\/\/schema.org","@graph":[{"@type":"Article","@id":"https:\/\/research.cchmc.org\/lungimage\/?p=19234#article","isPartOf":{"@id":"https:\/\/research.cchmc.org\/lungimage\/?p=19234"},"author":{"name":"kitc5i","@id":"https:\/\/research.cchmc.org\/lungimage\/#\/schema\/person\/94220f9638d78ff90fe4cd1d5743081f"},"headline":"PLISH In situ Hybridization of 200 \u00b5m Human Lung Tissue PACT for SFTPC","datePublished":"2018-02-20T20:14:41+00:00","dateModified":"2018-02-26T17:09:49+00:00","mainEntityOfPage":{"@id":"https:\/\/research.cchmc.org\/lungimage\/?p=19234"},"wordCount":724,"publisher":{"@id":"https:\/\/research.cchmc.org\/lungimage\/#organization"},"inLanguage":"en-US"},{"@type":"WebPage","@id":"https:\/\/research.cchmc.org\/lungimage\/?p=19234","url":"https:\/\/research.cchmc.org\/lungimage\/?p=19234","name":"PLISH In situ Hybridization of 200 \u00b5m Human Lung Tissue PACT for SFTPC -","isPartOf":{"@id":"https:\/\/research.cchmc.org\/lungimage\/#website"},"datePublished":"2018-02-20T20:14:41+00:00","dateModified":"2018-02-26T17:09:49+00:00","breadcrumb":{"@id":"https:\/\/research.cchmc.org\/lungimage\/?p=19234#breadcrumb"},"inLanguage":"en-US","potentialAction":[{"@type":"ReadAction","target":["https:\/\/research.cchmc.org\/lungimage\/?p=19234"]}]},{"@type":"BreadcrumbList","@id":"https:\/\/research.cchmc.org\/lungimage\/?p=19234#breadcrumb","itemListElement":[{"@type":"ListItem","position":1,"name":"Home","item":"https:\/\/research.cchmc.org\/lungimage\/"},{"@type":"ListItem","position":2,"name":"PLISH In situ Hybridization of 200 \u00b5m Human Lung Tissue PACT for SFTPC"}]},{"@type":"WebSite","@id":"https:\/\/research.cchmc.org\/lungimage\/#website","url":"https:\/\/research.cchmc.org\/lungimage\/","name":"","description":"Lung Images","publisher":{"@id":"https:\/\/research.cchmc.org\/lungimage\/#organization"},"potentialAction":[{"@type":"SearchAction","target":{"@type":"EntryPoint","urlTemplate":"https:\/\/research.cchmc.org\/lungimage\/?s={search_term_string}"},"query-input":{"@type":"PropertyValueSpecification","valueRequired":true,"valueName":"search_term_string"}}],"inLanguage":"en-US"},{"@type":"Organization","@id":"https:\/\/research.cchmc.org\/lungimage\/#organization","name":"Cincinnati Children's Hospital Medical Center","url":"https:\/\/research.cchmc.org\/lungimage\/","logo":{"@type":"ImageObject","inLanguage":"en-US","@id":"https:\/\/research.cchmc.org\/lungimage\/#\/schema\/logo\/image\/","url":"https:\/\/research.cchmc.org\/lungimage\/wp-content\/uploads\/2018\/05\/CCHMC-LOGO-II.png","contentUrl":"https:\/\/research.cchmc.org\/lungimage\/wp-content\/uploads\/2018\/05\/CCHMC-LOGO-II.png","width":668,"height":277,"caption":"Cincinnati Children's Hospital Medical Center"},"image":{"@id":"https:\/\/research.cchmc.org\/lungimage\/#\/schema\/logo\/image\/"}},{"@type":"Person","@id":"https:\/\/research.cchmc.org\/lungimage\/#\/schema\/person\/94220f9638d78ff90fe4cd1d5743081f","name":"kitc5i","image":{"@type":"ImageObject","inLanguage":"en-US","@id":"https:\/\/research.cchmc.org\/lungimage\/#\/schema\/person\/image\/","url":"https:\/\/secure.gravatar.com\/avatar\/e4bb6585a3f4dcb372eaf2f60ace1104aa6c71adfb2a1fa31533d31bb61f4034?s=96&d=mm&r=g","contentUrl":"https:\/\/secure.gravatar.com\/avatar\/e4bb6585a3f4dcb372eaf2f60ace1104aa6c71adfb2a1fa31533d31bb61f4034?s=96&d=mm&r=g","caption":"kitc5i"},"url":"https:\/\/research.cchmc.org\/lungimage\/?author=1"}]}},"_links":{"self":[{"href":"https:\/\/research.cchmc.org\/lungimage\/index.php?rest_route=\/wp\/v2\/posts\/19234","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/research.cchmc.org\/lungimage\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/research.cchmc.org\/lungimage\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/research.cchmc.org\/lungimage\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/research.cchmc.org\/lungimage\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=19234"}],"version-history":[{"count":9,"href":"https:\/\/research.cchmc.org\/lungimage\/index.php?rest_route=\/wp\/v2\/posts\/19234\/revisions"}],"predecessor-version":[{"id":19497,"href":"https:\/\/research.cchmc.org\/lungimage\/index.php?rest_route=\/wp\/v2\/posts\/19234\/revisions\/19497"}],"wp:attachment":[{"href":"https:\/\/research.cchmc.org\/lungimage\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=19234"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/research.cchmc.org\/lungimage\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=19234"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/research.cchmc.org\/lungimage\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=19234"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}