{"id":17181,"date":"2024-05-23T11:35:59","date_gmt":"2024-05-23T02:35:59","guid":{"rendered":"https:\/\/www.thermofisher.com\/blog\/learning-at-the-bench\/?p=17181"},"modified":"2024-09-12T18:23:08","modified_gmt":"2024-09-12T09:23:08","slug":"flow-cytometry-immunostaining-workshop-gsd-ts-tr-24028","status":"publish","type":"post","link":"https:\/\/www.thermofisher.com\/blog\/learning-at-the-bench\/flow-cytometry-immunostaining-workshop-gsd-ts-tr-24028\/","title":{"rendered":"\u3010\u3084\u3063\u3066\u307f\u305f\u3011\u6297\u4f53\u3092\u8cfc\u5165\u3057\u305f\u307e\u307e\u306e\u539f\u6db2\u3067\u53cd\u5fdc\u3055\u305b\u3066\u307f\u305f"},"content":{"rendered":"<p>\u6297\u4f53\u53cd\u5fdc\u3055\u305b\u3066\u3082\u30b7\u30b0\u30ca\u30eb\u304c\u4f4e\u304f\u3001\u3046\u307e\u304f\u691c\u51fa\u304c\u3067\u304d\u306a\u3044\u2026\u305d\u3093\u306a\u3068\u304d\u306f\u6297\u4f53\u91cf\u3092\u5897\u3084\u3057\u3066\u518d\u5ea6\u30c6\u30b9\u30c8\u3057\u305f\u304f\u306a\u308a\u307e\u3059\u3002\u6297\u4f53\u91cf\u306f\u591a\u3044\u65b9\u304c\u3088\u304f\u53cd\u5fdc\u3059\u308b\u306e\u306f\u5f53\u7136\u3067\u3059\u304c\u3001\u6975\u7aef\u306b\u591a\u304f\u3057\u305f\u5834\u5408\u3001\u3069\u3093\u306a\u30c7\u30fc\u30bf\u306b\u306a\u308b\u306e\u304b\u3002\u6c17\u306b\u306a\u3063\u305f\u306e\u3067\u3084\u3063\u3066\u307f\u307e\u3057\u305f\uff01<\/p>\n<h2>\u6750\u6599\u3068\u65b9\u6cd5<\/h2>\n<p>\u6750\u6599\uff1a<\/p>\n<ul>\n<li>\u30d2\u30c8\u5b50\u5bae\u9838\u304c\u3093\u7531\u6765\u7d30\u80de\u682aHeLa\uff082\u00d7105 cells\/sample\uff09<\/li>\n<li>CD44 Monoclonal Antibody (IM7), FITC\u3000#11-0441-82, 0.5 mg\/mL\uff08PBS, pH 7.2, 0.09% sodium azide\uff09<\/li>\n<li>Rat IgG2b kappa Isotype Control (eB149\/10H5), FITC, eBioscience\u3000#11-4031-82, 0.5 mg\/mL\uff08PBS, pH 7.2, 0.09% sodium azide\uff09<\/li>\n<li><a href=\"https:\/\/www.thermofisher.com\/jp\/ja\/home\/life-science\/cell-analysis\/flow-cytometry\/flow-cytometers\/attune-nxt-flow-cytometer.html?CID=gsd_pca_r04_jp_cp0000_pjt0000_gsd00000_0so_blg_flow_cytometry_immunostaining_workshop_gsd_ts_tr_24028_Social_LAB\" rel=\"noopener\" target=\"_blank\">Invitrogen\u2122 Attune\u2122 Nxt Flow Cytometer<\/a><\/li>\n<\/ul>\n<p>\u65b9\u6cd5\uff1a<\/p>\n<ol>\n<li>\u9060\u5fc3\u5206\u96e2\u3057\u3066\u4e0a\u6e05\u3092\u9664\u53bb\u3057\u305fHeLa\u7d30\u80de\u306e\u6c88\u6bbf\u306bFITC\u6a19\u8b58CD44\u6297\u4f53\u307e\u305f\u306fFITC\u6a19\u8b58Isotype control\u3092\u6dfb\u52a0\u3057\u3066\u61f8\u6fc1\u3002\u5168\u91cf\u306f100 \u03bcL\u306b\u306a\u308b\u3088\u3046\u306bPBS\u3068\u6df7\u5408\u3057\u3066\u6db2\u91cf\u3092\u8abf\u6574\u3057\u305f\u3002\u6297\u4f53\u30fbIsotype control\u306e\u6dfb\u52a0\u91cf\u306f\u4e0b\u8868\u306e\u3068\u304a\u308a\u3002<\/li>\n<li>\u5ba4\u6e29\u306730\u5206\u9593\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30c8<\/li>\n<li>\u9060\u5fc3\u5206\u96e2\u5f8c\u3001\u4e0a\u6e05\u3092\u9664\u53bb\u3057\u3066500 \u03bcL\u306ePBS\u306b\u61f8\u6fc1\u3057\u3001Attune NxT Flow Cytometer\u3067\u6e2c\u5b9a<\/li>\n<\/ol>\n<table>\n<caption><b>\u8868\uff0e\u4f7f\u7528\u3057\u305fCD44\u6297\u4f53\u3068Isotype Control\u306e\u91cf<\/b><\/caption>\n<tbody style=\"border-bottom: none\">\n<tr bgcolor=\"#6495ED\">\n<td style=\"text-align: center\" width=\"10\"><b><font color=\"white\">Sample\uff03<\/font><\/b><\/td>\n<td style=\"text-align: center\" width=\"10\"><b><font color=\"white\">\u6297\u4f53\u539f\u6db2<br \/>\uff08\u03bcL\uff09<\/font><\/b><\/td>\n<td style=\"text-align: center\" width=\"10\"><b><font color=\"white\">\u6297\u4f53\u91cf<br \/>\uff08\u03bcg\/test\uff09<\/font><\/b><\/td>\n<td style=\"text-align: center\" width=\"10\"><b><font color=\"white\">\u5099\u8003<\/font><\/b><\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" width=\"10\">1<\/td>\n<td style=\"text-align: center\" width=\"10\">100<\/td>\n<td style=\"text-align: center\" width=\"10\">50<\/td>\n<td style=\"text-align: center\" width=\"10\">\u539f\u6db2\u305d\u306e\u307e\u307e<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" width=\"10\">2<\/td>\n<td style=\"text-align: center\" width=\"10\">50<\/td>\n<td style=\"text-align: center\" width=\"10\">25<\/td>\n<td style=\"text-align: center\" width=\"10\"><\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" width=\"10\">3<\/td>\n<td style=\"text-align: center\" width=\"10\">20<\/td>\n<td style=\"text-align: center\" width=\"10\">10<\/td>\n<td style=\"text-align: center\" width=\"10\"><\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" width=\"10\">4<\/td>\n<td style=\"text-align: center\" width=\"10\">10<\/td>\n<td style=\"text-align: center\" width=\"10\">5<\/td>\n<td style=\"text-align: center\" width=\"10\"><\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" width=\"10\">5<\/td>\n<td style=\"text-align: center\" width=\"10\">5<\/td>\n<td style=\"text-align: center\" width=\"10\">2.5<\/td>\n<td style=\"text-align: center\" width=\"10\"><\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" width=\"10\">6<\/td>\n<td style=\"text-align: center\" width=\"10\">2.5<\/td>\n<td style=\"text-align: center\" width=\"10\">1.25<\/td>\n<td style=\"text-align: center\" width=\"10\"><\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" width=\"10\">7<\/td>\n<td style=\"text-align: center\" width=\"10\">1<\/td>\n<td style=\"text-align: center\" width=\"10\">0.5<\/td>\n<td style=\"text-align: center\" width=\"10\">\u30e1\u30fc\u30ab\u30fc\u63a8\u5968\u91cf<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" width=\"10\">8<\/td>\n<td style=\"text-align: center\" width=\"10\">0.5<\/td>\n<td style=\"text-align: center\" width=\"10\">0.25<\/td>\n<td style=\"text-align: center\" width=\"10\"><\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" width=\"10\">9<\/td>\n<td style=\"text-align: center\" width=\"10\">0.25<\/td>\n<td style=\"text-align: center\" width=\"10\">0.125<\/td>\n<td style=\"text-align: center\" width=\"10\"><\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" width=\"10\">10<\/td>\n<td style=\"text-align: center\" width=\"10\">0.125<\/td>\n<td style=\"text-align: center\" width=\"10\">0.063<\/td>\n<td style=\"text-align: center\" width=\"10\"><\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" width=\"10\">11<\/td>\n<td style=\"text-align: center\" width=\"10\">0.0625<\/td>\n<td style=\"text-align: center\" width=\"10\">0.031<\/td>\n<td style=\"text-align: center\" width=\"10\"><\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" width=\"10\">12<\/td>\n<td style=\"text-align: center\" width=\"10\">0.03125<\/td>\n<td style=\"text-align: center\" width=\"10\">0.016<\/td>\n<td style=\"text-align: center\" width=\"10\"><\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" width=\"10\">13<\/td>\n<td style=\"text-align: center\" width=\"10\">0.015<\/td>\n<td style=\"text-align: center\" width=\"10\">0.08<\/td>\n<td style=\"text-align: center\" width=\"10\"><\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" width=\"10\">14<\/td>\n<td style=\"text-align: center\" width=\"10\">0<\/td>\n<td style=\"text-align: center\" width=\"10\">0<\/td>\n<td style=\"text-align: center\" width=\"10\">Negative Control<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: left\" width=\"50\" colspan=\"4\">\u203bIsotype Control\u306f\u3001\u5bfe\u8c61\u306e\u6297\u4f53\u3068\u540c\u3058\u30a2\u30a4\u30bd\u30bf\u30a4\u30d7\u3067\u540c\u3058\u86cd\u5149\u6a19\u8b58\u304c\u3055\u308c\u305f\u975e\u7279\u7570\u7684\u306a\u30a4\u30e0\u30ce\u30b0\u30ed\u30d6\u30ea\u30f3\u6eb6\u6db2\uff08\u4eca\u56de\u306fFITC\u6a19\u8b58\u3055\u308c\u305fRat IgG2b\uff09\u3002\u6297\u4f53\u306a\u3057\u306e\u30cd\u30ac\u30c6\u30a3\u30d6\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u306b\u6bd4\u3079\u3001\u3088\u308a\u6b63\u78ba\u306b\u30d0\u30c3\u30af\u30b0\u30e9\u30a6\u30f3\u30c9\u3092\u53cd\u6620\u3057\u305f\u30cd\u30ac\u30c6\u30a3\u30d6\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u3068\u3057\u3066\u7528\u3044\u3089\u308c\u308b\u3002<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<h2>\u7d50\u679c<\/h2>\n<p>\u307e\u305a\u306f\u30d2\u30b9\u30c8\u30b0\u30e9\u30e0\u30d7\u30ed\u30c3\u30c8\u3092\u3054\u89a7\u304f\u3060\u3055\u3044\uff08\u56f31\uff09\u3002<br \/>\nCD44\u6297\u4f53\u3067\u53cd\u5fdc\u3055\u305b\u305f\u30b5\u30f3\u30d7\u30eb\u306f\u3001\u6297\u4f53\u539f\u6db2\u3067\u53cd\u5fdc\u3055\u305b\u305f\u3082\u306e\u3068\u3001\u63a8\u5968\u91cf\u3067\u53cd\u5fdc\u3055\u305b\u305f\u3082\u306e\u3067\u307b\u3068\u3093\u3069\u9055\u3044\u306f\u898b\u3089\u308c\u307e\u305b\u3093\u3067\u3057\u305f\u3002\u4e00\u65b9\u3001Isotype Control\u3067\u306f\u63a8\u5968\u91cf\u3067\u306f\u6297\u4f53\u306a\u3057\uff08\u30cd\u30ac\u30c6\u30a3\u30d6\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\uff1aData not shown\uff09\u3068\u540c\u7b49\u306e\u30d7\u30ed\u30c3\u30c8\u3067\u3057\u305f\u304c\u3001\u6297\u4f53\u539f\u6db2\u3067\u306f\u307e\u308b\u3067\u967d\u6027\u30b5\u30f3\u30d7\u30eb\u3068\u898b\u7d1b\u3046\u3070\u304b\u308a\u306e\u30d7\u30ed\u30c3\u30c8\u3068\u306a\u308a\u307e\u3057\u305f\u3002\u3082\u3061\u308d\u3093\u3053\u308c\u306f\u30d0\u30c3\u30af\u30b0\u30e9\u30a6\u30f3\u30c9\u304c\u4e0a\u304c\u3063\u3066\u3044\u308b\u305f\u3081\u3067\u3001\u3084\u306f\u308a\u6297\u4f53\u539f\u6db2\u3067\u306f\u6fc3\u3059\u304e\u308b\u3053\u3068\u304c\u5206\u304b\u308a\u307e\u3057\u305f\u3002<\/p>\n<div id=\"attachment_17185\" style=\"width: 659px\" class=\"wp-caption aligncenter\"><img loading=\"lazy\" decoding=\"async\" aria-describedby=\"caption-attachment-17185\" src=\"http:\/\/admin.acceleratingscience.com\/learning-at-the-bench\/wp-content\/uploads\/sites\/13\/2024\/05\/684da3d9d959a96c7329031e07e64c45.jpg\" alt=\"\u56f31. \u30d2\u30b9\u30c8\u30b0\u30e9\u30e0\u30d7\u30ed\u30c3\u30c8\n\u6297\u4f53\u539f\u6db2\u3067\u53cd\u5fdc\uff08\u4e0a\u6bb5\uff09\u3001\u63a8\u5968\u91cf\u3067\u53cd\u5fdc\uff08\u4e0b\u6bb5\uff09\" width=\"649\" height=\"613\" class=\"size-full wp-image-17185\" srcset=\"https:\/\/admin.acceleratingscience.com\/learning-at-the-bench\/wp-content\/uploads\/sites\/13\/2024\/05\/684da3d9d959a96c7329031e07e64c45.jpg 649w, https:\/\/admin.acceleratingscience.com\/learning-at-the-bench\/wp-content\/uploads\/sites\/13\/2024\/05\/684da3d9d959a96c7329031e07e64c45-300x283.jpg 300w\" sizes=\"auto, (max-width: 649px) 100vw, 649px\" \/><p id=\"caption-attachment-17185\" class=\"wp-caption-text\">\u56f31. \u30d2\u30b9\u30c8\u30b0\u30e9\u30e0\u30d7\u30ed\u30c3\u30c8<br \/>\u6297\u4f53\u539f\u6db2\u3067\u53cd\u5fdc\uff08\u4e0a\u6bb5\uff09\u3001\u63a8\u5968\u91cf\u3067\u53cd\u5fdc\uff08\u4e0b\u6bb5\uff09<\/p><\/div>\n<p>\u305d\u308c\u3067\u306f\u3001\u6297\u4f53\u306f\u3069\u306e\u304f\u3089\u3044\u6fc3\u304f\u3057\u3066\u3082\u3088\u3044\u306e\u3067\u3057\u3087\u3046\u304b\u3002\u56f32\u3067\u306f\u6297\u4f53\u3092\u539f\u6db2\u304b\u3089\u7d046700\u500d\u307e\u3067\u6bb5\u968e\u7684\u306b\u8584\u3081\u3066\u53cd\u5fdc\u3055\u305b\u3001\u30d5\u30ed\u30fc\u30b5\u30a4\u30c8\u30e1\u30c8\u30ea\u30fc\u3067\u5f97\u3089\u308c\u305f\u30b7\u30b0\u30ca\u30eb\u5f37\u5ea6\uff08MFI: Mean of Fluorescence Intensity\uff09\u3092\u30b0\u30e9\u30d5\u306b\u3057\u305f\u3082\u306e\u3067\u3059\u3002<\/p>\n<div id=\"attachment_17186\" style=\"width: 932px\" class=\"wp-caption aligncenter\"><img loading=\"lazy\" decoding=\"async\" aria-describedby=\"caption-attachment-17186\" src=\"http:\/\/admin.acceleratingscience.com\/learning-at-the-bench\/wp-content\/uploads\/sites\/13\/2024\/05\/8b73ac454b2691bb8cef4a2d6aea3700.jpg\" alt=\"\u56f32. \u5404\u6297\u4f53\u91cf\u3067\u53cd\u5fdc\u3055\u305b\u305f\u30b7\u30b0\u30ca\u30eb\u5f37\u5ea6\nMFI\u5024\u306e\u30d7\u30ed\u30c3\u30c8\u56f3\uff08\u4e0a\u6bb5\uff09\u3068CD44\u6297\u4f53\u3068Isotype Control\u306eMFI\u6bd4\uff08\u4e0b\u6bb5\uff09\" width=\"922\" height=\"613\" class=\"size-full wp-image-17186\" srcset=\"https:\/\/admin.acceleratingscience.com\/learning-at-the-bench\/wp-content\/uploads\/sites\/13\/2024\/05\/8b73ac454b2691bb8cef4a2d6aea3700.jpg 922w, https:\/\/admin.acceleratingscience.com\/learning-at-the-bench\/wp-content\/uploads\/sites\/13\/2024\/05\/8b73ac454b2691bb8cef4a2d6aea3700-300x199.jpg 300w, https:\/\/admin.acceleratingscience.com\/learning-at-the-bench\/wp-content\/uploads\/sites\/13\/2024\/05\/8b73ac454b2691bb8cef4a2d6aea3700-768x511.jpg 768w, https:\/\/admin.acceleratingscience.com\/learning-at-the-bench\/wp-content\/uploads\/sites\/13\/2024\/05\/8b73ac454b2691bb8cef4a2d6aea3700-96x65.jpg 96w\" sizes=\"auto, (max-width: 922px) 100vw, 922px\" \/><p id=\"caption-attachment-17186\" class=\"wp-caption-text\">\u56f32. \u5404\u6297\u4f53\u91cf\u3067\u53cd\u5fdc\u3055\u305b\u305f\u30b7\u30b0\u30ca\u30eb\u5f37\u5ea6<br \/>MFI\u5024\u306e\u30d7\u30ed\u30c3\u30c8\u56f3\uff08\u4e0a\u6bb5\uff09\u3068CD44\u6297\u4f53\u3068Isotype Control\u306eMFI\u6bd4\uff08\u4e0b\u6bb5\uff09<\/p><\/div>\n<p>Isotype Control\u3067\u306f\u63a8\u5968\u91cf\u306e5\u500d\uff085 \u03bcL\/test\uff09\u307e\u3067\u306f\u30d0\u30c3\u30af\u30b0\u30e9\u30f3\u30c9\u5024\u3082\u307b\u307c\u5909\u308f\u3089\u306a\u3044\u3053\u3068\u304c\u5206\u304b\u308a\u307e\u3057\u305f\u3002\u4e00\u65b9\u3001CD44\u6297\u4f53\u306f\u63a8\u5968\u91cf\u3067\u53cd\u5fdc\u3055\u305b\u3055\u305f\u30b5\u30f3\u30d7\u30eb\u306b\u6bd4\u3079\u30015\u500d\u91cf\u306e\u5834\u5408\u306f\u30b7\u30b0\u30ca\u30eb\u5024\u304c\u4e0a\u6607\u3057\u3066\u3044\u308b\u3088\u3046\u3067\u3059\u304c\u3001\u305d\u306e\u5dee\u306f\u308f\u305a\u304b\u3067\u3057\u305f\u3002<br \/>\nCD44\u3068Isotype Control\u306e\u30b7\u30b0\u30ca\u30eb\u5024\u306e\u6bd4\u3001\u3044\u308f\u3086\u308bSN\u6bd4\uff08\u56f32\u4e0b\u6bb5\uff09\u3092\u898b\u308b\u3068\u3001\u63a8\u5968\u91cf\u3067\u3042\u308b1 \u03bcL\/test\u3092\u542b\u30800.5\uff5e5 \u03bcL\/test\u306e\u7bc4\u56f2\u3067\u304a\u304a\u3080\u306d\u30d4\u30fc\u30af\u306b\u9054\u3057\u3066\u3044\u307e\u3057\u305f\u3002\u3064\u307e\u308a\u4eca\u56de\u306e\u5b9f\u9a13\u7cfb\u3067\u306f\u3001\u3053\u306e\u7bc4\u56f2\u304c\u9069\u3057\u305f\u6297\u4f53\u91cf\u3068\u3044\u3046\u3053\u3068\u304c\u5206\u304b\u308a\u307e\u3057\u305f\u3002<br \/>\n\u306a\u304a\u6297\u4f53\u304c\u5c11\u306a\u304f\u3066\u3082\u3001\u4f8b\u3048\u3070\u63a8\u5968\u91cf\u306e10\u5206\u306e1\u7a0b\u5ea6\u3067\u3082Isotype Control\u306b\u6bd4\u3079\u3066\u660e\u77ad\u306b\u30d2\u30b9\u30c8\u30b0\u30e9\u30e0\u4e0a\u3067CD44\u967d\u6027\u3067\u3042\u308b\u3053\u3068\u304c\u5206\u304b\u308a\u307e\u3057\u305f\uff08Data not shown\uff09\u3002\u5b9a\u91cf\u3067\u306f\u306a\u304f\u3001\u967d\u6027\u304b\u9670\u6027\u304b\u5206\u304b\u308c\u3070\u3088\u3044\u5834\u5408\u306f\u7bc0\u7d04\u306e\u305f\u3081\u306b\u8584\u3081\u308b\u3053\u3068\u3082\u3067\u304d\u305d\u3046\u3067\u3059\u3002<br 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