{"id":281,"date":"2023-07-07T12:44:14","date_gmt":"2023-07-07T04:44:14","guid":{"rendered":"http:\/\/yuhongtaolab.com\/?post_type=resource&#038;p=281"},"modified":"2023-07-07T12:44:14","modified_gmt":"2023-07-07T04:44:14","slug":"hesc-facs-sorting-protocol","status":"publish","type":"resource","link":"http:\/\/yuhongtaolab.com\/en\/resource\/hesc-facs-sorting-protocol\/","title":{"rendered":"hESC FACS sorting protocol"},"content":{"rendered":"<p>1. wash hESC once with DMEM\/F12 to remove most dead cells or cell debris.<br \/>\n2. add 1ml\/well accutase, incubate at 37\u2103 for 8-10min.<br \/>\n3. Gently triturate cell aggregates with P1000 tips for 5-6 times to get homogeneous single cell suspension.<br \/>\n4. transfer cell suspension into 15ml tube, centrifuge at 200g for 5min.<br \/>\n5. Discard supernatant, resuspend cell pellets in 1-2ml mTeSR1 (Y27632+).<br \/>\n6. Filter cell suspension through 40um strainer into 15ml tube.<br \/>\n7. Count cell density, adjust density if needed.<br \/>\n8. Place cell suspension on ice before loading onto FACS instrument.<br \/>\n9. Load cell suspension into FACS sorter, collect target cells into mTeSR1 (Y27632+).<br \/>\n10. after FACS, collect cell pellets by centrifuge, resuspend cell pellets in mTeSR1 (Y27632+)<\/p>","protected":false},"featured_media":0,"template":"","class_list":["post-281","resource","type-resource","status-publish","hentry"],"acf":[],"_links":{"self":[{"href":"http:\/\/yuhongtaolab.com\/en\/wp-json\/wp\/v2\/resource\/281","targetHints":{"allow":["GET"]}}],"collection":[{"href":"http:\/\/yuhongtaolab.com\/en\/wp-json\/wp\/v2\/resource"}],"about":[{"href":"http:\/\/yuhongtaolab.com\/en\/wp-json\/wp\/v2\/types\/resource"}],"wp:attachment":[{"href":"http:\/\/yuhongtaolab.com\/en\/wp-json\/wp\/v2\/media?parent=281"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}