{"id":1,"date":"2025-01-24T23:21:16","date_gmt":"2025-01-24T15:21:16","guid":{"rendered":"https:\/\/tiand.top\/?p=1"},"modified":"2025-01-28T09:16:48","modified_gmt":"2025-01-28T01:16:48","slug":"hello-world","status":"publish","type":"post","link":"https:\/\/tiand.top\/index.php\/2025\/01\/24\/hello-world\/","title":{"rendered":"\u5206\u79bb\u5c0f\u9f20\u80a0\u9053T\u7ec6\u80de\uff08IV\u578b\u80f6\u539f\u9176\uff09"},"content":{"rendered":"\n

\u3010\u5b9e\u9a8c\u6750\u6599\u3011<\/strong><\/p>\n\n\n\n

    \n
  1. 40% Percoll\u6eb6\u6db2\uff1a<\/strong> Percoll\u5de5\u4f5c\u6db2<\/strong> + 1\u00d7HBSS\uff084:6\u7a00\u91ca\uff09 Percoll<\/strong> **\u5de5\u4f5c\u6db2\uff1a**Percoll\u539f\u6db2+10\u00d7HBSS\uff089:1\u7a00\u91ca\uff09<\/li>\n\n\n\n
  2. \u9884\u6d88\u5316\u6db2<\/strong>\uff084<\/strong> mL\/colon\uff0c\u542b10 mM EDTA, 1 mM DTT\u7684\u65e0Ca2+\/Mg2+\u7684HBSS\u6eb6\u6db2\uff09<\/li>\n\n\n\n
  3. \u6d88\u5316\u6db2<\/strong>\uff084<\/strong> mL\/colon\uff0c\u542b4% FBS\uff0c10 mM HEPES\uff0c0.5 mg\/ml Collagenase IV\uff0c0.2 mg\/ml DNase I\u7684RPMI 1640\u6eb6\u6db2\uff09<\/li>\n\n\n\n
  4. 50X<\/strong> EDTA\uff1a 0.5M\u50a8\u5b58\u6db2\uff0cHBSS\u7a00\u91ca\uff08Sigma, Cat# EDS-100G\uff09<\/li>\n\n\n\n
  5. 1000X<\/strong> DTT\uff08DL-Dithiothreitol solution\uff09\uff1a 1M\u539f\u6db2 \uff08Sigma, Cat# 43816-10ML\uff09<\/li>\n\n\n\n
  6. 100X<\/strong> HEPES\uff1a\u50a8\u5b58\u6db21 M (Thermo, Cat# 15630080)<\/li>\n\n\n\n
  7. 100X<\/strong> Collagenase IV\uff1a\u50a8\u5b58\u6db250 mg\/ml\uff0c100mg+2mL HBSS\u7a00\u91ca100uL\/tube \uff08Sigma, C5138\uff09<\/li>\n\n\n\n
  8. 100X<\/strong> DNase I\uff1a\u50a8\u5b58\u6db220 mg\/ml\uff0c40mg+2mL 1640\u7a00\u91ca100uL\/tube (Sigma, Cat# DN25)<\/li>\n<\/ol>\n\n\n\n

    <\/p>\n\n\n\n

    \u3010\u5b9e\u9a8c\u65b9\u6cd5\u3011<\/strong><\/p>\n\n\n\n

      \n
    1. \u53d6\u6b63\u5e38B6\u5c0f\u9f20\u5c0f\u80a0\u7ec4\u7ec7\uff0c\u4ed4\u7ec6\u53bb\u9664\u7cfb\u819c\u7ec4\u7ec7\uff0c\u7f6e\u4e8e\u51b0\u9884\u51b7DPBS\u4e2d\uff1b<\/li>\n\n\n\n
    2. \u7528\u773c\u79d1\u954a\u5b50\u5939\u4f4f\u80a0\u9053\u4e00\u7aef\uff0c\u7528\u6ce8\u5c04\u5668+\u526a\u88c1\u7684200\u03bcL\u67aa\u5934\u5438\u53d6DPBS\u63d2\u5165\u80a0\u7ba1\u4e00\u7aef\uff0c\u51b2\u51fa\u80a0\u5185\u7caa\u4fbf\uff1b<\/li>\n\n\n\n
    3. \u526a\u53bb\u80a0\u9053\u8868\u9762\u8102\u80aa\u7ec4\u7ec7\u548cPayer Patch\uff1b<\/li>\n\n\n\n
    4. \u7eb5\u5411\u526a\u5f00\u80a0\u9053\uff0c\u5728\u51b0\u9884\u51b7DPBS\u6dae\u6d17\u6570\u6b21\uff0c\u81f3\u65e0\u7d6e\u72b6\u7269\u8131\u843d\uff0c\u526a\u62101cm\u957f\u7684\u5c0f\u6bb5\u5907\u7528\uff1b<\/li>\n\n\n\n
    5. \u80a0\u6bb5\u52a0\u51654 mL\u9884\u6d88\u5316\u6db2<\/strong>\u4e2d\uff0c37\u2103\u5b75\u80b220min\uff0c\u6c34\u5e73\u9707\u8361\uff08130<\/strong>\u901f\uff09\uff1b<\/li>\n\n\n\n
    6. \u5c06\u80a0\u6bb5\u548c\u9884\u6d88\u5316\u6db2\u7ecf\u8fc770 \u03bcm\u7b5b\u7f51\u8fc7\u6ee4\uff0c\u6d41\u51fa\u6db2\u5373\u4e3aIELs<\/strong>\uff0c\u7ec6\u80de\u8ba1\u6570\uff1b<\/li>\n\n\n\n
    7. \u5c06\u80a0\u6bb5\u7ecfPBS\u6dae\u6d17(\u6b64\u6b65\u9aa4\u91cd\u8981<\/strong>)\u540e\uff0c\u526a\u4e3a2~4mm\u7247\u6bb5\uff1b<\/li>\n\n\n\n
    8. \u5c06\u5c0f\u80a0\u6bb5\u7ec4\u7ec7\u52a0\u51654 mL\u6d88\u5316\u6db2<\/strong>\uff0c37\u2103\u5b75\u80b230 min\uff0c\u6c34\u5e73\u7f13\u6162\u6447\u6643**\uff0890<\/strong>\u901f\uff09**\u6da1\u65cb20s\uff1b<\/li>\n\n\n\n
    9. \u52a0\u51653\u500d\u4f53\u79ef\u51b0\u9884\u51b7MACS\u7ec8\u6b62\u6d88\u5316\uff1b<\/li>\n\n\n\n
    10. \u6df7\u5408\u6db2\u4f53\u572870 \u03bcm\u7b5b\u7f51\u4e0a\u8fc7\u6ee4\uff08\u5269\u4f59\u5757\u72b6\u7ec4\u7ec7\u8f83\u591a\u65f6\u53ef\u7814\u78e8\uff09\uff0c\u6536\u96c6\u6d41\u51fa\u7ec6\u80de\uff1b<\/li>\n\n\n\n
    11. \u79bb\u5fc3500g 10min 20\u2103\uff0c\u5f03\u4e0a\u6e05\uff1b<\/li>\n\n\n\n
    12. \u6c89\u6dc0\u91cd\u60ac\u4e8e10 mL 40% Percoll\u6eb6\u6db2\uff1b<\/li>\n\n\n\n
    13. \u79bb\u5fc31000g\uff0c30 min\uff0c\u53471\u964d1\uff0c20\u2103\uff0c\u5438\u53bb\u8868\u5c42\u6742\u8d28\u53ca\u4e0a\u6e05\uff0c\u91cd\u60ac\u6c89\u6dc0\uff1b<\/li>\n\n\n\n
    14. \u52a0\u516510 mL PBS\uff0c\u8fc7\u4e00\u6b2170uM\u7b5b\u7f51\uff0c\u79bb\u5fc3\u6d17\u7ec6\u80de400g 5min<\/li>\n\n\n\n
    15. \u6d41\u5f0f\u68c0\u6d4b\u7ec6\u80de\u6c89\u6dc0\u4e2d\u7684\u6dcb\u5df4\u548c\u9ad3\u7cfb\u7ec6\u80de\u6bd4\u4f8b\u3002<\/li>\n<\/ol>\n","protected":false},"excerpt":{"rendered":"

      \u3010\u5b9e\u9a8c\u6750\u6599\u3011 \u3010\u5b9e\u9a8c\u65b9\u6cd5\u3011<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_jetpack_memberships_contains_paid_content":false,"footnotes":""},"categories":[8],"tags":[],"class_list":["post-1","post","type-post","status-publish","format-standard","hentry","category-protocols"],"jetpack_featured_media_url":"","jetpack_sharing_enabled":true,"_links":{"self":[{"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/posts\/1","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/comments?post=1"}],"version-history":[{"count":4,"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/posts\/1\/revisions"}],"predecessor-version":[{"id":297,"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/posts\/1\/revisions\/297"}],"wp:attachment":[{"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/media?parent=1"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/categories?post=1"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/tags?post=1"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}