\nunder the direction of Elisabeth ERRAZURIZ-CERDA<\/strong><\/a> from CIQLE<\/strong> (Lyon Est Quantitative Imaging Center).<\/strong><\/a><\/p>\n
<\/p>\n
EM stain biological samples (Hepatocytes, Thrombocytes) using an O.T.E-based solution before inclusion on cell pellets.<\/h4>\nprotocol:<\/strong><\/h4>\n\n- \u00a0Prior preparation of OTE at 0.2%<\/strong><\/a> (solution previously aliquoted and frozen)
\n\u2022 1% glutaraldehyde fixation in 0.1M Na cacodylate buffer, pH 7.4 2h at 4 degrees
\n\u2022 3 rinses in 0.1M Na cacodylate for a few hours to 1 night at +4\u00b0C
\n\u2022 Post fixation: Osmium tetroxide 1%+1.5% potassium ferrocyanide in cacodylate buffer in 0.1M cacodylate buffer, pH 7.4.
\n\u2022 Rinses in 0.1M Na cacodylate buffer, pH 7.4
\n\u2022 Contrast OTE 0.2% in cacodylate 1h TA
\n\u2022 Rinses with distilled water.
\n\u2022 Dehydration: ethanol 30\u00b0 (5min), 50\u00b0(5min), 70\u00b0(5min), 95\u00b0(5min), 100\u00b0(3x10min)
\n\u2022 Substitution: Epon resin 1v. \/ Absolute ethanol 1v.
\n\u2022 Pure Epon resin impregnation under vacuum
\n\u2022 Inclusion with Epon +DMP30. Polymerization at 60\u00b0 for 48 hours
\n\u2022 Contrast of ultrafine sections 5min with lead citrate
\n\u2022 Rinse with water<\/p>\nConclusion:<\/strong><\/h3>\nthis contrast EM Stain agent based on Oolong Tea Extract (O.T.E)<\/strong><\/a> allows a beautiful visualization of intracytoplasmic constituents and represents a good substitute for uranyl acetate.<\/li>\n<\/ul>\n\n- Our contrast experience with the use of OTE on two tissues types : Hepatocyte\u00a0 liver and thrombocyte tissue is very conclusive with good visualization of the different cellular compartments and organelles. OTE \u201csafe stain\u201d allowed us to rule out toxic contrastants such as uranyl acetate.
\nResults :<\/h3>\n
Photo1: Visualization of contacts between mitochondria and reticulum in a hepatocyte<\/span>.<\/li>\n![\"\"](\"https:\/\/www.em-grade.com\/wp-content\/uploads\/2024\/03\/OTE-hepatocyte-Cicle-lyon-150x150.jpg\")
<\/a><\/li>\n- \u2022 Gly: Glycogen grains, Mit: Mitochondria, REG: Rough endoplasmic reticulum, Lip: Lipid droplet
\n\u2022 Photo credit: Jennifer Rieusset- CARMEN \u2013 INSERM U1060 \u2013 Lyon<\/a> and CIQLE<\/a> (Lyon Est Quantitative Imaging Center)
\n\u2022 Equipment: JEOL EUROPE<\/a> JEM 1400 transmission electron microscope, Gatan Inc<\/a>. Orius600 camera<\/li>\n- <\/li>\n
- click on image to enlarge the photo.<\/span><\/li>\n
- <\/li>\n<\/ul>\n
<\/p>\n
<\/p>\n
\n- Photo2: Counting alpha granules in patient’s blood platelets.<\/span><\/li>\n
![\"\"](\"https:\/\/www.em-grade.com\/wp-content\/uploads\/2024\/03\/OTE-hepatocyte-CiQle-lyon-150x150.jpg\")
<\/a>
\n\u2022 DTS: Dense tubular system, Gly: Glycogen, OCS: Open canalicular system, \u03bb-G: Alpha granules
\n\u2022 Photo credit: Ana\u00eblle De Wreede- Hospices Civils de Lyon \u2013 HCL<\/a> \u2013 Hemostasis Laboratory and CIQLE (Lyon Est Quantitative Imaging Center)
\n\u2022 Equipment: JEOL EUROPE<\/a> JEM 1400 transmission electron microscope, Gatan Inc<\/a>. Orius600 camera<\/li>\n- <\/li>\n
- <\/li>\n
- click on image to enlarge the photo.<\/span><\/li>\n<\/ul>\n","protected":false},"excerpt":{"rendered":"
Customer feedback usingEM Stain : OTE (Oolong Tea Extract). We share with you an experience of using a non-toxic EM stain like OTE, under the direction of Elisabeth ERRAZURIZ-CERDA from…<\/p>\n","protected":false},"author":8,"featured_media":8801,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"_seopress_titles_title":"","_seopress_titles_desc":"","_seopress_robots_index":"","_seopress_robots_follow":"","_seopress_robots_imageindex":"","_seopress_robots_snippet":"","_seopress_robots_primary_cat":"","_seopress_robots_breadcrumbs":"","_seopress_robots_freeze_modified_date":"","_seopress_robots_custom_modified_date":"","_seopress_robots_canonical":"","_seopress_social_fb_title":"","_seopress_social_fb_desc":"","_seopress_social_fb_img":"","_seopress_social_fb_img_attachment_id":0,"_seopress_social_fb_img_width":0,"_seopress_social_fb_img_height":0,"_seopress_social_twitter_title":"","_seopress_social_twitter_desc":"","_seopress_social_twitter_img":"","_seopress_social_twitter_img_attachment_id":0,"_seopress_social_twitter_img_width":0,"_seopress_social_twitter_img_height":0,"_seopress_redirections_value":"","_seopress_redirections_enabled":"","_seopress_redirections_enabled_regex":"","_seopress_redirections_logged_status":"","_seopress_redirections_param":"","_seopress_redirections_type":0,"_seopress_analysis_target_kw":"","_seopress_news_disabled":"","_seopress_video_disabled":"","_seopress_video":[],"_seopress_pro_schemas_manual":[],"_seopress_pro_rich_snippets_disable_all":"","_seopress_pro_rich_snippets_disable":[],"_seopress_pro_schemas":[],"itsec_x_frame_options":"","footnotes":""},"categories":[782],"tags":[798],"class_list":["post-8799","post","type-post","status-publish","format-standard","has-post-thumbnail","category-em-stain-it","tag-ote-it"],"acf":[],"_links":{"self":[{"href":"https:\/\/www.em-grade.com\/it\/wp-json\/wp\/v2\/posts\/8799","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.em-grade.com\/it\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.em-grade.com\/it\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.em-grade.com\/it\/wp-json\/wp\/v2\/users\/8"}],"replies":[{"embeddable":true,"href":"https:\/\/www.em-grade.com\/it\/wp-json\/wp\/v2\/comments?post=8799"}],"version-history":[{"count":0,"href":"https:\/\/www.em-grade.com\/it\/wp-json\/wp\/v2\/posts\/8799\/revisions"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.em-grade.com\/it\/wp-json\/wp\/v2\/media\/8801"}],"wp:attachment":[{"href":"https:\/\/www.em-grade.com\/it\/wp-json\/wp\/v2\/media?parent=8799"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.em-grade.com\/it\/wp-json\/wp\/v2\/categories?post=8799"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.em-grade.com\/it\/wp-json\/wp\/v2\/tags?post=8799"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}
- \n
- \u00a0Prior preparation of OTE at 0.2%<\/strong><\/a> (solution previously aliquoted and frozen)
\n\u2022 1% glutaraldehyde fixation in 0.1M Na cacodylate buffer, pH 7.4 2h at 4 degrees
\n\u2022 3 rinses in 0.1M Na cacodylate for a few hours to 1 night at +4\u00b0C
\n\u2022 Post fixation: Osmium tetroxide 1%+1.5% potassium ferrocyanide in cacodylate buffer in 0.1M cacodylate buffer, pH 7.4.
\n\u2022 Rinses in 0.1M Na cacodylate buffer, pH 7.4
\n\u2022 Contrast OTE 0.2% in cacodylate 1h TA
\n\u2022 Rinses with distilled water.
\n\u2022 Dehydration: ethanol 30\u00b0 (5min), 50\u00b0(5min), 70\u00b0(5min), 95\u00b0(5min), 100\u00b0(3x10min)
\n\u2022 Substitution: Epon resin 1v. \/ Absolute ethanol 1v.
\n\u2022 Pure Epon resin impregnation under vacuum
\n\u2022 Inclusion with Epon +DMP30. Polymerization at 60\u00b0 for 48 hours
\n\u2022 Contrast of ultrafine sections 5min with lead citrate
\n\u2022 Rinse with water<\/p>\nConclusion:<\/strong><\/h3>\n
this contrast EM Stain agent based on Oolong Tea Extract (O.T.E)<\/strong><\/a> allows a beautiful visualization of intracytoplasmic constituents and represents a good substitute for uranyl acetate.<\/li>\n<\/ul>\n
- \n
- Our contrast experience with the use of OTE on two tissues types : Hepatocyte\u00a0 liver and thrombocyte tissue is very conclusive with good visualization of the different cellular compartments and organelles. OTE \u201csafe stain\u201d allowed us to rule out toxic contrastants such as uranyl acetate.
\nResults :<\/h3>\n
Photo1: Visualization of contacts between mitochondria and reticulum in a hepatocyte<\/span>.<\/li>\n
<\/a><\/li>\n- \u2022 Gly: Glycogen grains, Mit: Mitochondria, REG: Rough endoplasmic reticulum, Lip: Lipid droplet
\n\u2022 Photo credit: Jennifer Rieusset- CARMEN \u2013 INSERM U1060 \u2013 Lyon<\/a> and CIQLE<\/a> (Lyon Est Quantitative Imaging Center)
\n\u2022 Equipment: JEOL EUROPE<\/a> JEM 1400 transmission electron microscope, Gatan Inc<\/a>. Orius600 camera<\/li>\n- <\/li>\n
- click on image to enlarge the photo.<\/span><\/li>\n
- <\/li>\n<\/ul>\n
<\/p>\n
<\/p>\n
- \n
- Photo2: Counting alpha granules in patient’s blood platelets.<\/span><\/li>\n<\/a>
\n\u2022 DTS: Dense tubular system, Gly: Glycogen, OCS: Open canalicular system, \u03bb-G: Alpha granules
\n\u2022 Photo credit: Ana\u00eblle De Wreede- Hospices Civils de Lyon \u2013 HCL<\/a> \u2013 Hemostasis Laboratory and CIQLE (Lyon Est Quantitative Imaging Center)
\n\u2022 Equipment: JEOL EUROPE<\/a> JEM 1400 transmission electron microscope, Gatan Inc<\/a>. Orius600 camera<\/li>\n- <\/li>\n
- <\/li>\n
- click on image to enlarge the photo.<\/span><\/li>\n<\/ul>\n","protected":false},"excerpt":{"rendered":"
Customer feedback usingEM Stain : OTE (Oolong Tea Extract). We share with you an experience of using a non-toxic EM stain like OTE, under the direction of Elisabeth ERRAZURIZ-CERDA from…<\/p>\n","protected":false},"author":8,"featured_media":8801,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"_seopress_titles_title":"","_seopress_titles_desc":"","_seopress_robots_index":"","_seopress_robots_follow":"","_seopress_robots_imageindex":"","_seopress_robots_snippet":"","_seopress_robots_primary_cat":"","_seopress_robots_breadcrumbs":"","_seopress_robots_freeze_modified_date":"","_seopress_robots_custom_modified_date":"","_seopress_robots_canonical":"","_seopress_social_fb_title":"","_seopress_social_fb_desc":"","_seopress_social_fb_img":"","_seopress_social_fb_img_attachment_id":0,"_seopress_social_fb_img_width":0,"_seopress_social_fb_img_height":0,"_seopress_social_twitter_title":"","_seopress_social_twitter_desc":"","_seopress_social_twitter_img":"","_seopress_social_twitter_img_attachment_id":0,"_seopress_social_twitter_img_width":0,"_seopress_social_twitter_img_height":0,"_seopress_redirections_value":"","_seopress_redirections_enabled":"","_seopress_redirections_enabled_regex":"","_seopress_redirections_logged_status":"","_seopress_redirections_param":"","_seopress_redirections_type":0,"_seopress_analysis_target_kw":"","_seopress_news_disabled":"","_seopress_video_disabled":"","_seopress_video":[],"_seopress_pro_schemas_manual":[],"_seopress_pro_rich_snippets_disable_all":"","_seopress_pro_rich_snippets_disable":[],"_seopress_pro_schemas":[],"itsec_x_frame_options":"","footnotes":""},"categories":[782],"tags":[798],"class_list":["post-8799","post","type-post","status-publish","format-standard","has-post-thumbnail","category-em-stain-it","tag-ote-it"],"acf":[],"_links":{"self":[{"href":"https:\/\/www.em-grade.com\/it\/wp-json\/wp\/v2\/posts\/8799","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.em-grade.com\/it\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.em-grade.com\/it\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.em-grade.com\/it\/wp-json\/wp\/v2\/users\/8"}],"replies":[{"embeddable":true,"href":"https:\/\/www.em-grade.com\/it\/wp-json\/wp\/v2\/comments?post=8799"}],"version-history":[{"count":0,"href":"https:\/\/www.em-grade.com\/it\/wp-json\/wp\/v2\/posts\/8799\/revisions"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.em-grade.com\/it\/wp-json\/wp\/v2\/media\/8801"}],"wp:attachment":[{"href":"https:\/\/www.em-grade.com\/it\/wp-json\/wp\/v2\/media?parent=8799"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.em-grade.com\/it\/wp-json\/wp\/v2\/categories?post=8799"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.em-grade.com\/it\/wp-json\/wp\/v2\/tags?post=8799"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}
- Our contrast experience with the use of OTE on two tissues types : Hepatocyte\u00a0 liver and thrombocyte tissue is very conclusive with good visualization of the different cellular compartments and organelles. OTE \u201csafe stain\u201d allowed us to rule out toxic contrastants such as uranyl acetate.