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树突状细胞亚群检测操作程序

树突状细胞亚群检测操作程序

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Published by vic1116

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Published by: vic1116 on Mar 25, 2009
Copyright:Attribution Non-commercial

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02/01/2013

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\u5357\u4eac\u533b\u79d1\u5927\u5b66\u9644\u5c5e\u5357\u4eac\u7b2c\u4e00\u533b\u9662
\u4e2d\u5fc3\u5b9e\u9a8c\u5ba4
Central Laboratory Of Nanjing First
Hospital Affiliated To Nanjing
Medical University
\u6811\u7a81\u72b6\u7ec6\u80de\u4e9a\u7fa4\u68c0\u6d4b\u64cd
\u4f5c\u7a0b\u5e8f
\u7f16 \u53f7\uff1aSop-CL-FCM-010
\u65e5 \u671f\uff1a2009-03-05
\u7248 \u672c\uff1a\u7b2c\u4e00\u7248
\u7b2c1 \u9875
\u51712 \u9875
\u4e00\u3001\u76ee\u7684\uff1a
\u4fdd\u8bc1\u6d41\u5f0f\u7ec6\u80de\u4eea\u68c0\u6d4b\u5916\u5468\u8840\u6216\u9aa8\u9ad3\u6db2\u4e2d\u6811\u7a81\u72b6\u7ec6\u80de\u4e9a\u7fa4\u7684\u5de5\u4f5c\u8d28\u91cf\uff0c\u65e8\u5728
\u4fdd\u8bc1\u6811\u7a81\u72b6\u7ec6\u80de\u4e9a\u7fa4\u68c0\u6d4b\u6b63\u786e\u7684\u64cd\u4f5c\u7a0b\u5e8f\uff0c\u4ee5\u63a7\u5236\u68c0\u6d4b\u7ed3\u679c\u7684\u7cbe\u786e\u5ea6\u3002
\u4e8c\u3001\u4fee\u6539\u7a0b\u5e8f\uff1a
\u672c\u6807\u51c6\u64cd\u4f5c\u7a0b\u5e8f\u7684\u6539\u52a8\uff0c\u53ef\u7531\u4efb\u4e00\u4f7f\u7528\u672cSOP \u7684\u5de5\u4f5c\u4eba\u5458\u63d0\u51fa\uff0c\u5e76
\u62a5\u7ecf\u4e0b\u8ff0\u4eba\u5458\u6279\u51c6\u7b7e\u5b57\uff1a\u8d28\u91cf\u4e3b\u7ba1\u3001\u6388\u6743\u4eba\uff08\u79d1\u4e3b\u4efb\uff09\uff0c\u65b9\u53ef\u6539\u52a8\u3002
\u4e09\u3001\u9002\u7528\u8303\u56f4\uff1a
\u5916\u5468\u8840\u6216\u9aa8\u9ad3\u6db2\u4e2d\u6811\u7a81\u72b6\u7ec6\u80de\u4e9a\u7fa4\u68c0\u6d4b\u3002
\u56db\u3001 \u6eaf\u6e90\uff1a
\u7f8e\u56fd BD \u516c\u53f8\u4ea7\u54c1\u8bf4\u660e\u4e66\uff08CAT:340546\uff09\u3002
\u4e94\u3001\u5b9e\u9a8c\u539f\u7406\uff1a

\u4eba\u5916\u5468\u8840\u4e2d\u6811\u7a81\u72b6\u7ec6\u80de\u4e3b\u8981\u662f\u4e0d\u6210\u719f DC\uff0c\u6709\u4e24\u4e2a\u4e9a\u7fa4\uff0c\u4e0e\u8840\u6db2\u4e2d\u5176\u5b83\u7ec6\u80de \u76f8\u6bd4\uff0cDC \u7ec6\u80de\u6709\u72ec\u7279\u7684\u514d\u75ab\u8868\u578b\u3002\u4ee5\u6b64\u4e3a\u4f9d\u636e\uff0c\u6765\u68c0\u6d4b\u5916\u5468\u8840\u4e2d\u6811\u7a81\u72b6\u7ec6\u80de\u7684 \u767e\u5206\u542b\u91cf\u3002DC1 \u4e0d\u8868\u8fbe lin\uff08CD19\u3001CD20\u3001CD56\u3001CD14\u3001CD3\uff09\uff0c\u9ad8\u8868\u8fbe HLA-DR \u548c CD11c\uff1bDC2 \u4e0d\u8868\u8fbe lin\uff08CD19\u3001CD20\u3001CD56\u3001CD14\u3001CD3\uff09\uff0c\u9ad8\u8868\u8fbe HLA-DR \u548c CD123\u3002\u5229\u7528\u56db\u8272\u6d41\u5f0f\u7ec6\u80de\u4eea\u5206\u6790\u6280\u672f\uff0c\u68c0\u6d4b\u51fa DC1 \u4e0e DC2 \u7684\u767e\u5206\u6bd4\u3002

\u516d\u3001\u4e3b\u8981\u8bd5\u5242\uff1a
1\u3001Lineage Cocktail-FITC (\u7f8e\u56fd BD Biosciences \u516c\u53f8\uff1bCAT:340546)\uff0c
2\u3001Mouse IgG1-PE (\u7f8e\u56fd BD Biosciences \u516c\u53f8\uff1bCAT:349043)
3\u3001Mouse IgG1-APC (\u7f8e\u56fd BD Biosciences \u516c\u53f8\uff1bCAT:340442)
4\u3001CD123-PE(\u7f8e\u56fd BD Biosciences \u516c\u53f8\uff1bCAT:340545)

5\u3001CD11c-APC(\u7f8e\u56fd BD Biosciences \u516c\u53f8\uff1bCAT:340544)
6\u3001HLA-DR-PreCP(\u7f8e\u56fd BD Biosciences \u516c\u53f8\uff1bCAT:347364)
\u4e03\u3001\u5b9e\u9a8c\u64cd\u4f5c\uff1a
1\u3001\u6807\u672c\u91c7\u96c6\uff1a\u4f7f\u7528 EDTA-K2\u6297\u51dd\u771f\u7a7a\u91c7\u8840\u7ba1\uff0c\u62bd\u53d6\u9759\u8109\u8840 2ml\uff1b\u6216\u65b0\u9c9c\u809d\u7d20

\u6297\u51dd\u9aa8\u9ad3 1.0ml\u3002
2\u3001\u67d3\u8272\u548c\u56fa\u5b9a\u7ec6\u80de\uff1a
\uff081\uff09\u6807\u672c\u7ba1\u7f16\u53f7 A\uff0c\u53e6\u53d6\u6d41\u5f0f\u4e13\u7528\u8bd5\u7ba1\u4e24\u53ea AT(\u6d4b\u5b9a)\u3001AC\uff08\u5bf9\u7167\uff09\u3002
\uff082\uff09AT \u7ba1\u52a0\u5165 Lineage Cocktail-FITC\u3001HLA-DR-PreCP\u3001CD123-
PE\u3001CD11c-APC \u5404 20ul\u3002AC \u7ba1\u52a0\u5165 Lineage Cocktail-FITC\u3001HLA-DR-
PreCP\u3001Mouse IgG1-PE \u548c Mouse IgG1-APC \u5404 20ul\u3002

\uff083\uff09AT\u3001AC \u7ba1\u4e2d\u52a0\u5165\u5168\u8840\u6216\u9aa8\u9ad3 100ul\uff0c\u6df7\u5747\u3002
\uff084\uff09\u7f6e\u5ba4\u6e29\uff0c\u907f\u5149\u5b75\u80b2 15min\u3002
\uff085\uff09\u52a0\u5165 FACSLysing \u6eb6\u8840\u6db2 2.0ml\uff0c\u7f6e\u5ba4\u6e29\uff0c\u907f\u5149\u5b75\u80b2 10min,\u5bf9\u5149\u89c2\u5bdf

\u7ea2\u7ec6\u80de\u6eb6\u89e3\u662f\u5426\u5b8c\u5168\uff0c\u5f85\u6eb6\u6db2\u900f\u660e\u3002
\uff086\uff09300g \u79bb\u5fc3 5min\uff0c\u5f03\u4e0a\u6e05\u6db2\uff0c\u6d17\u6da4\u4e24\u6b21\u3002
3\u3001\u6df7\u5300\u540e\u4e0a\u673a\u68c0\u6d4b\uff1b\u5206\u6790\u7ed3\u679c\uff1b\u6253\u5370\u5b9e\u9a8c\u62a5\u544a\u3002
\u516b\u3001\u53c2\u8003\u503c\uff1a
DC1:0.28%\u00b10.07%\uff1bDC2:0.22%\u00b10.09%
\u4e5d\u3001\u4e34\u5e8a\u610f\u4e49\uff1a
\u6811\u7a81\u72b6\u7ec6\u80de\uff08dendritic cells\uff0cDC\uff09\u662f\u7531\u7f8e\u56fd\u5b66\u8005\u4e8e 1973\u5e74\u9996\u5148\u53d1
\u73b0\u3002\u5b83\u662f\u76ee\u524d\u6240\u77e5\u7684\u673a\u4f53\u5185\u6297\u539f\u9012\u5448\u529f\u80fd\u6700\u5f3a\u7684\u6297\u539f\u9012\u5448\u7ec6\u80de\uff0c\u4e5f\u662f\u552f\u4e00
\u80fd\u5c06\u6297\u539f \u9012\u5448\u7ed9\u521d\u59cbT \u7ec6\u80de\uff0c \u6fc0\u53d1\u521d\u6b21\u514d\u75ab \u5e94\u7b54\u7684 APC\uff0c \u56e0\u6b64\uff0c\u5728\u514d\u75ab \u5e94
\u7b54\u7684\u8bf1\u5bfc\u4e2d\u5177\u6709\u7279\u6b8a\u7684\u5730\u4f4d\u3002DC1/DC2 \u7684\u8bf1\u5bfc\u5206\u5316\u76f4\u63a5\u5f71\u54cdTH1/TH2 \u7684\u5e73\u8861\uff0c
\u4eba\u4eec\u53ef\u4ee5\u901a\u8fc7\u8c03\u8282 DC1/DC2 \u6765\u5f71\u54cd TH1/TH2 \u7684\u5e73\u8861\u3002TH1\u8fc7\u5ea6\u589e\u9ad8\u901a\u5e38\u5f15\u8d77
\u591a\u53d1\u6027\u786c\u5316\u75c5\u7b49\u81ea\u8eab\u514d\u75ab\u75be\u75c5\uff0cTH2 \u8fc7\u9ad8\uff0c\u5e38\u5e38\u4e0e\u54ee\u5598\u3001\u8fc7\u654f\u7b49\u75be\u75c5\u76f8\u5173\u3002

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