{"id":1641,"date":"2026-07-27T09:53:05","date_gmt":"2026-07-27T09:53:05","guid":{"rendered":"https:\/\/www.hxicollege.com\/?p=1641"},"modified":"2026-07-27T18:37:55","modified_gmt":"2026-07-27T10:37:55","slug":"customer-case-ecnu-huang-zhengqi-phalloidin-actin-1641","status":"publish","type":"post","link":"https:\/\/www.hxicollege.com\/index.php\/2026\/07\/27\/customer-case-ecnu-huang-zhengqi-phalloidin-actin-1641\/","title":{"rendered":"20230418-\u534e\u4e1c\u5e08\u8303-\u9ec4\u6b63\u9f50\u8001\u5e08\u6837\u54c1\u68c0\u6d4b\u62a5\u544a"},"content":{"rendered":"<style>\n  \/* ===== \u6574\u7bc7\u5bb9\u5668 ===== *\/\n  .hx-article{\n    max-width:1080px;\n    margin:0 auto;\n    padding:40px 28px 64px;\n    font-family:-apple-system,BlinkMacSystemFont,\"Segoe UI\",\"PingFang SC\",\"Hiragino Sans GB\",\"Microsoft YaHei\",sans-serif;\n    color:#1f2328;\n    line-height:1.85;\n    font-size:16px;\n  }\n  \/* ===== \u6848\u4f8b\u6807\u9898 ===== *\/\n  .hx-article h2{\n    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.hx-block .info-line{color:#cbd5e1;}\n  body.aft-dark-mode .hx-block .info-line b{color:#f1f5f9;}\n  body.aft-dark-mode .hx-specs .item{background:#0f172a;color:#cbd5e1;border-color:#3b82f6;}\n  body.aft-dark-mode .hx-specs .item b{color:#f1f5f9;}\n  body.aft-dark-mode .hx-raw{background:#0f172a;border-color:#334155;color:#94a3b8;}\n  body.aft-dark-mode .hx-raw:hover{background:#1e293b;color:#f1f5f9;}\n  body.aft-dark-mode .hx-end{color:#64748b;border-color:#334155;}\n  \/* ===== \u79fb\u52a8\u7aef\u56de\u9000 ===== *\/\n  @media (max-width:760px){\n    .hx-grid{flex-direction:column;gap:14px;}\n    .hx-protocol{padding:16px;}\n    .hx-figure img{max-height:none;}\n    .hx-fig-grid.cols-2,\n    .hx-fig-grid.cols-3,\n    .hx-fig-grid.cols-4{grid-template-columns:repeat(2,1fr);}\n    .hx-specs{grid-template-columns:1fr;}\n  }\n<\/style>\n<div class=\"hx-article\">\n<p>  <!-- ===== \u6848\u4f8b\u6807\u9898 ===== --><\/p>\n<h2>\u6848\u4f8b 03\uff1a\u534e\u4e1c\u5e08\u8303\u5927\u5b66-\u9ec4\u6b63\u9f50\u8001\u5e08\u6837\u54c1\u68c0\u6d4b\u62a5\u544a\uff08\u9b3c\u7b14\u73af\u80bd\u00b7\u808c\u52a8\u86cb\u767d\uff09<\/h2>\n<p>  <!-- ===== \u9879\u76ee\u6982\u51b5 hero ===== --><\/p>\n<div class=\"hx-hero\">\n<h3>\ud83d\udccb \u9879\u76ee\u6982\u51b5<\/h3>\n<p><b>\u5ba2\u6237\u5355\u4f4d\uff1a<\/b>\u534e\u4e1c\u5e08\u8303\u5927\u5b66<\/p>\n<p><b>\u9879\u76ee\u8d1f\u8d23\u4eba\uff1a<\/b>\u9ec4\u6b63\u9f50\u8001\u5e08<\/p>\n<p><b>\u9879\u76ee\u76ee\u6807\uff1a<\/b>\u9b3c\u7b14\u73af\u80bd\uff08Phalloidin\uff09\u808c\u52a8\u86cb\u767d\uff08Actin\uff09\u8367\u5149\u6807\u8bb0\u6837\u54c1\u7684\u591a\u5e73\u53f0\u68c0\u6d4b\u8bc4\u4f30\uff0c\u8986\u76d6 <b>Acti-Stain 555 Phalloidin<\/b> \u4e0e <b>Alexa Fluor\u2122 532 Phalloidin<\/b> \u4e24\u6b3e\u67d3\u6599\uff0c\u8de8 2023 \u5e74 4 \u6708\u81f3 8 \u6708\u5171 6 \u8f6e\u68c0\u6d4b\u8fed\u4ee3\u3002<\/p>\n<p><b>\u9879\u76ee\u6587\u6863\uff1a<\/b><a href=\"https:\/\/vsen1hqljj.feishu.cn\/docx\/CPowdJUEio6wLrxR2dBc8BWHnng\" target=\"_blank\" rel=\"noopener\">\u534e\u4e1c\u5e08\u8303\u5927\u5b66-\u9ec4\u6b63\u9f50\u8001\u5e08\u5b9a\u5236\u9879\u76ee\uff08\u98de\u4e66\uff09 \u2197<\/a><\/p>\n<\/p><\/div>\n<p>  <!-- ===== \u6807\u8bb0\u539f\u6599\u8bb0\u5f55 ===== --><\/p>\n<div class=\"hx-section\">\n<h3><span class=\"badge\">Reagents<\/span>\u6807\u8bb0\u539f\u6599\u8bb0\u5f55<\/h3>\n<div class=\"hx-block\">\n<h4>\ud83c\udfa8 Acti-Stain 555 Phalloidin<\/h4>\n<p>\u6765\u6e90\uff1a<a href=\"https:\/\/www.cytoskeleton.com\/phdh1\" target=\"_blank\" rel=\"noopener\">cytoskeleton.com\/phdh1<\/a><\/p>\n<\/p><\/div>\n<div class=\"hx-block\">\n<h4>\ud83c\udfa8 Alexa Fluor\u2122 532 \u9b3c\u7b14\u73af\u80bd<\/h4>\n<p><b>\u6fc0\u53d1\/\u53d1\u5c04\u6ce2\u957f\uff1a<\/b>531 \/ 554 nm<\/p>\n<p>\u4ea7\u54c1\u94fe\u63a5\uff1a<a href=\"https:\/\/www.thermofisher.cn\/order\/catalog\/product\/A22282\" target=\"_blank\" rel=\"noopener\">thermofisher.cn\/order\/catalog\/product\/A22282<\/a><\/p>\n<div class=\"hx-fig-grid cols-2\">\n<figure class=\"hx-figure\">\n          <img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/screenshot_thermo_532.webp\" alt=\"Thermo Fisher Alexa Fluor 532 Phalloidin \u4ea7\u54c1\u9875\"><figcaption>Thermo Fisher \u4ea7\u54c1\u9875\uff08\u9b3c\u7b14\u73af\u80bd Alexa 532\uff09<\/figcaption><\/figure>\n<div class=\"hx-block\">\n<p><b>In DMSO\uff1a<\/b>150 \u03bcL 66 \u03bcM stock<\/p>\n<p><b>In methanol\uff1a<\/b>1.5 mL 6.6 \u03bcM stock<\/p>\n<p class=\"info-line\">\u672c\u9879\u76ee\u9009\u62e9 <b>\u7532\u9187\u6eb6\u89e3<\/b>\uff0c\u540e\u7eed\u9700\u505a\u6d53\u5ea6\u6ef4\u5b9a\uff08titration\uff09\u4ee5\u5e73\u8861\u4fe1\u53f7\u5f3a\u5ea6\u4e0e\u80cc\u666f\u8367\u5149\u3002<\/p>\n<\/p><\/div>\n<\/p><\/div>\n<p>\u4f7f\u7528\u8bf4\u660e PDF\uff1a<a href=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/MAN0001777_Phalloidins_UG.pdf\" target=\"_blank\" rel=\"noopener\">MAN0001777_Phalloidins_UG.pdf \u2197<\/a><\/p>\n<\/p><\/div>\n<\/p><\/div>\n<p>  <!-- ===== \u68c0\u6d4b\u8bbe\u5907 ===== --><\/p>\n<div class=\"hx-section\">\n<h3><span class=\"badge\">Equipment<\/span>\u68c0\u6d4b\u8bbe\u5907\uff1aOlympus FV3000<\/h3>\n<p><b>60\u00d7 \u7269\u955c LSM \u6fc0\u5149\u5149\u6e90\uff1a<\/b><\/p>\n<div class=\"hx-specs\">\n<div class=\"item\">405 nm \u00b7 <b>50 mW<\/b><\/div>\n<div class=\"item\">445 nm \u00b7 <b>75 mW<\/b><\/div>\n<div class=\"item\">488 nm \u00b7 <b>20 mW<\/b><\/div>\n<div class=\"item\">514 nm \u00b7 <b>40 mW<\/b><\/div>\n<div class=\"item\">561 nm \u00b7 <b>20 mW<\/b><\/div>\n<div class=\"item\">594 nm \u00b7 <b>20 mW<\/b><\/div>\n<div class=\"item\">640 nm \u00b7 <b>40 mW<\/b><\/div>\n<\/p><\/div>\n<\/p><\/div>\n<p>  <!-- ===== 20230418 Actin-Alexa555 ===== --><\/p>\n<div class=\"hx-section\">\n<h3><span class=\"badge\">20230418<\/span>Actin-Alexa555 \u6837\u54c1\u68c0\u6d4b\u8be6\u60c5<\/h3>\n<p><b>\u5bf9\u6bd4\u6837\u54c1\uff1a<\/b>\u4e24\u6279 BSC-1 \u7ec6\u80de\uff0c\u5206\u522b\u4f7f\u7528 <b>4% PFA<\/b> \u4e0e <b>4% PFA + 1% GA<\/b> \u4e24\u79cd\u56fa\u5b9a\u65b9\u6848\uff1b\u6fc0\u5149\u4ec5\u5f00 0.3%\uff0c\u8bc4\u4f30\u4f4e\u6fc0\u5149\u4e0b\u4e24\u79cd\u56fa\u5b9a\u5242\u7684\u7ec6\u8282\u4fdd\u7559\u80fd\u529b\u3002<\/p>\n<table class=\"hx-table\">\n<thead>\n<tr>\n<th style=\"width:18%\">\u7ef4\u5ea6<\/th>\n<th>\u6837\u54c1 1\uff08S01\uff09<\/th>\n<th>\u6837\u54c1 2\uff08S02\uff09<\/th>\n<\/tr>\n<\/thead>\n<tbody>\n<tr>\n<td><b>\u6807\u7b7e<\/b><\/td>\n<td>S01-Alexa555 phalloidin<br \/>4% PFA Fix<br \/>Bright\u3001lowBG \u2014 \u6837\u54c1 A<\/td>\n<td>S02-Alexa555 phalloidin<br \/>4% PFA + 1% GA Fix<br \/>Bright\u3001higher BG\u3001good details \u2014 \u6837\u54c1 B<\/td>\n<\/tr>\n<tr>\n<td><b>\u56fa\u5b9a\u5242<\/b><\/td>\n<td>4% PFA<\/td>\n<td>4% PFA + 1% GA<\/td>\n<\/tr>\n<tr>\n<td><b>\u6807\u8bb0\u4fe1\u606f<\/b><\/td>\n<td>Alexa555 phalloidin<\/td>\n<td>Alexa555 phalloidin<\/td>\n<\/tr>\n<tr>\n<td><b>\u6837\u54c1\u5907\u6ce8<\/b><\/td>\n<td>\u80cc\u666f\u66f4\u5e72\u51c0\uff0c\u96fe\u72b6\u4e0e\u4e1d\u72b6\u7ec6\u8282\u8db3\u591f\uff0c\u6837\u54c1\u975e\u5e38\u660e\u4eae\uff080.3% \u6fc0\u5149\uff09<\/td>\n<td>\u80cc\u666f\u8f83\u9ad8\uff0c\u4f46\u4fe1\u53f7\u66f4\u4eae\uff0c\u96fe\u72b6\u4e0e\u4e1d\u72b6\u7ec6\u8282\u4fdd\u5b58\u66f4\u591a\uff080.3% \u6fc0\u5149\uff09<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<h4>60\u00d7 \u5171\u805a\u7126\u6210\u50cf\uff08S01 vs S02\uff09<\/h4>\n<div class=\"hx-fig-grid cols-3\">\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230418-S01-C01-561.webp\" alt=\"S01 C01\"><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230418-S01-C02-561.webp\" alt=\"S01 C02\"><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230418-S01-C03-561.webp\" alt=\"S01 C03\"><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230418-S02-C01-561.webp\" alt=\"S02 C01\"><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230418-S02-C02-561.webp\" alt=\"S02 C02\"><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230418-S02-C03-561.webp\" alt=\"S02 C03\"><\/figure>\n<\/p><\/div>\n<h4>20\u00d7 \u4f4e\u500d\u6982\u89c8<\/h4>\n<div class=\"hx-fig-grid cols-2\">\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230418-S01-C01-561-20x.webp\" alt=\"S01 20x\"><figcaption>S01 \u00b7 4% PFA \u00b7 20\u00d7<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230418-S02-C01-561-20x.webp\" alt=\"S02 20x\"><figcaption>S02 \u00b7 4% PFA + 1% GA \u00b7 20\u00d7<\/figcaption><\/figure>\n<\/p><\/div>\n<\/p><\/div>\n<p>  <!-- ===== Time scale ===== --><\/p>\n<div class=\"hx-section\">\n<h3><span class=\"badge\">Time<\/span>\u626b\u63cf\u65f6\u95f4\u5c3a\u5ea6<\/h3>\n<div class=\"hx-grid\">\n<div class=\"hx-col\">\n<figure class=\"hx-figure\">\n          <img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/time_sacle.webp\" alt=\"\u626b\u63cf\u65f6\u95f4\u5c3a\u5ea6\"><figcaption>\u5355\u50cf\u7d20 ~2 \u03bcs \u00b7 1024 \u00d7 1024 pixel \u00b7 \u626b\u63cf 3.217 s<\/figcaption><\/figure>\n<\/p><\/div>\n<div class=\"hx-col\">\n<div class=\"hx-block\">\n<p>\u5728 <b>2 \u03bcs\/pixel<\/b> \u7684\u626b\u63cf\u53c2\u6570\u4e0b\uff0c\u6574\u5f20 1024 \u00d7 1024 \u50cf\u7d20\u56fe\u91c7\u96c6\u8017\u65f6\u7ea6 <b>3.2 s<\/b>\u3002\u8be5\u53c2\u6570\u517c\u987e\u4e86\u6837\u54c1\u7684\u5149\u6f02\u767d\u63a7\u5236\u4e0e\u5fae\u7ba1\u7ec6\u8282\u7684\u6e05\u6670\u5ea6\uff0c\u662f\u540e\u7eed\u591a\u8f6e\u68c0\u6d4b\u8fed\u4ee3\u7684\u57fa\u7840\u8bbe\u7f6e\u3002<\/p>\n<\/p><\/div>\n<\/p><\/div>\n<\/p><\/div>\n<\/p><\/div>\n<p>  <!-- ===== 20230601 ===== --><\/p>\n<div class=\"hx-section\">\n<h3><span class=\"badge\">20230601<\/span>Actin-Alexa532 \u6837\u54c1\u68c0\u6d4b\u8be6\u60c5<\/h3>\n<h4>\u6807\u8bb0\u539f\u6599<\/h4>\n<figure class=\"hx-figure\">\n      <img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/image_2.webp\" alt=\"20230601 \u6807\u8bb0\u539f\u6599\"><br \/>\n    <\/figure>\n<h4>\u6210\u50cf\u68c0\u6d4b\u7ed3\u679c\uff08FV3000 60\u00d7\uff09<\/h4>\n<table class=\"hx-table\">\n<thead>\n<tr>\n<th>\u6837\u54c1<\/th>\n<th>S01<\/th>\n<th>S02<\/th>\n<th>S03<\/th>\n<th>S04<\/th>\n<\/tr>\n<\/thead>\n<tbody>\n<tr>\n<td><b>\u67d3\u6599<\/b><\/td>\n<td>Alexa Fluor 532 phalloidin<\/td>\n<td>Alexa Fluor 532 phalloidin<\/td>\n<td>Alexa Fluor 532 phalloidin<\/td>\n<td>Alexa Fluor 532 phalloidin<\/td>\n<\/tr>\n<tr>\n<td><b>\u6d53\u5ea6<\/b><\/td>\n<td>4%<\/td>\n<td>4%<\/td>\n<td>50%<\/td>\n<td>16.6%<\/td>\n<\/tr>\n<tr>\n<td><b>\u5b75\u80b2<\/b><\/td>\n<td>4 \u2103 \u8fc7\u591c\uff080602\uff09<\/td>\n<td>4 \u2103 \u8fc7\u591c\uff080602\uff09<\/td>\n<td>4 \u2103 \u8fc7\u591c\uff080602\uff09<\/td>\n<td>4 \u2103 \u8fc7\u591c\uff080602\uff09<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<div class=\"hx-fig-grid cols-4\">\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/202306002-S01-C01-60X-514.webp\" alt=\"S01 60X\"><figcaption>S01 \u00b7 4%<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/202306002-S02-C01-60X-514.webp\" alt=\"S02 60X\"><figcaption>S02 \u00b7 4%<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/202306012-S03-C01-60X-514.webp\" alt=\"S03 60X\"><figcaption>S03 \u00b7 50%<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/202306012-S04-C01-60X-514.webp\" alt=\"S04 60X\"><figcaption>S04 \u00b7 16.6%<\/figcaption><\/figure>\n<\/p><\/div>\n<h4>\u57fa\u6069\u58eb\u5bf9\u7167\uff0860\u00d7\uff09<\/h4>\n<div class=\"hx-fig-grid cols-3\">\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230603_3T3_JS01-C01-60X_GFP_7s7.webp\" alt=\"S01 \u57fa\u6069\u58eb GFP 7s\"><figcaption>S01 \u00b7 GFP \u00b7 7s<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230603_3T3_JS03-C01-60X_TXRED_6s.webp\" alt=\"S03 \u57fa\u6069\u58eb TXRED 6s\"><figcaption>S03 \u00b7 TXRED \u00b7 6s<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230603_3T3_JS04-C01-60X_TXRED_4.5s.webp\" alt=\"S04 \u57fa\u6069\u58eb TXRED 4.5s\"><figcaption>S04 \u00b7 TXRED \u00b7 4.5s<\/figcaption><\/figure>\n<\/p><\/div>\n<\/p><\/div>\n<p>  <!-- ===== 20230602 ===== --><\/p>\n<div class=\"hx-section\">\n<h3><span class=\"badge\">20230602<\/span>Actin-Alexa532 \u6837\u54c1\u68c0\u6d4b\u8be6\u60c5<\/h3>\n<h4>\u6807\u8bb0\u539f\u6599<\/h4>\n<figure class=\"hx-figure\">\n      <img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/image_4.webp\" alt=\"20230602 \u6807\u8bb0\u539f\u6599\"><br \/>\n    <\/figure>\n<h4>\u6210\u50cf\u68c0\u6d4b\u7ed3\u679c<\/h4>\n<table class=\"hx-table\">\n<thead>\n<tr>\n<th>\u6837\u54c1<\/th>\n<th>S01<\/th>\n<th>S02<\/th>\n<th>S03<\/th>\n<th>S04<\/th>\n<th>S05\uff08+ \u6807\u51c6\u6837\uff09<\/th>\n<\/tr>\n<\/thead>\n<tbody>\n<tr>\n<td><b>\u67d3\u6599<\/b><\/td>\n<td>Alexa Fluor 532<\/td>\n<td>Alexa Fluor 532<\/td>\n<td>Alexa Fluor 532<\/td>\n<td>Alexa Fluor 532<\/td>\n<td>Alexa Fluor 532<\/td>\n<\/tr>\n<tr>\n<td><b>\u6d53\u5ea6<\/b><\/td>\n<td>10%<\/td>\n<td>50%<\/td>\n<td>20%<\/td>\n<td>100%<\/td>\n<td>\u2014<\/td>\n<\/tr>\n<tr>\n<td><b>\u67d3\u8272\u65f6\u95f4<\/b><\/td>\n<td>20230602<\/td>\n<td>20230602<\/td>\n<td>20230602<\/td>\n<td>20230602<\/td>\n<td>20230602<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<h4>\u57fa\u6069\u58eb\u521d\u626b\uff08Brightness:1 \/ 6s\uff09<\/h4>\n<div class=\"hx-fig-grid cols-4\">\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/image_7.webp\" alt=\"S01 1\/6s\"><figcaption>S01 \u00b7 10%<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/image.webp\" alt=\"S02 1\/6s\"><figcaption>S02 \u00b7 50%<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230602_NIH3T3_S03_5X_-C01-60X.webp\" alt=\"S03 NIH3T3\"><figcaption>S03 \u00b7 20%<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/image_1.webp\" alt=\"S04 1\/6s\"><figcaption>S04 \u00b7 100%<\/figcaption><\/figure>\n<\/p><\/div>\n<div class=\"hx-callout\">\n      \ud83d\udcac <b>\u6d53\u5ea6\u8ba8\u8bba\uff1a<\/b>S02\uff0850% = 1:2\uff09\u7591\u4f3c\u7a00\u91ca\u504f\u7a00\uff1b\u5efa\u8bae\u5c06\u7532\u9187\u6eb6\u89e3\u7684\u67d3\u6599\u5148\u53d6 50 \u03bcL \u667e\u5e72\u4e3a\u7c89\u672b\uff0c\u91cd\u65b0\u6eb6\u4e8e 5 \u03bcL DMSO\uff0c\u5236\u5907 10\u00d7 \u6d53\u5ea6\u7684 Stock\u3002\n    <\/div>\n<h4>\u57fa\u6069\u58eb\u590d\u6d4b\uff08Brightness:1 \/ 12s \u00b7 4 \u2103 \u8fc7\u591c\u5b75\u80b2\uff09<\/h4>\n<div class=\"hx-fig-grid cols-4\">\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230603_3T3_S01-C02-60X_TXRED_12s.webp\" alt=\"S01 1\/12s\"><figcaption>S01 \u00b7 12s<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230603_3T3_S02-C02-60X_TXRED_12s.webp\" alt=\"S02 1\/12s\"><figcaption>S02 \u00b7 12s<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230603_3T3_S03-C02-60X_TEXRED_3s.webp\" alt=\"S03 1\/3s\"><figcaption>S03 \u00b7 3s \u00b7 Bright 1\/3s<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/image_3.webp\" alt=\"S04 1\/12s\"><figcaption>S04 \u00b7 12s<\/figcaption><\/figure>\n<\/p><\/div>\n<h4>\u6b63\u5bf9\u7167\u6837\u54c1\uff08S05\uff09<\/h4>\n<div class=\"hx-fig-grid cols-2\">\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/2023-06-03_CH2-01.webp\" alt=\"CH2-01\"><figcaption>CH2-01<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/2023-06-03_CH2-02.webp\" alt=\"CH2-02\"><figcaption>CH2-02<\/figcaption><\/figure>\n<\/p><\/div>\n<table class=\"hx-table\">\n<thead>\n<tr>\n<th colspan=\"2\">Positive control\uff08\u57fa\u6069\u58eb\uff09<\/th>\n<\/tr>\n<\/thead>\n<tbody>\n<tr>\n<td><b>\u66dd\u5149\u65f6\u95f4<\/b><\/td>\n<td>1 \/ 10 s<\/td>\n<\/tr>\n<tr>\n<td><b>\u901a\u9053<\/b><\/td>\n<td>CH2<\/td>\n<\/tr>\n<tr>\n<td><b>Gain<\/b><\/td>\n<td>+6 dB<\/td>\n<\/tr>\n<tr>\n<td><b>\u6fc0\u5149\u5f3a\u5ea6<\/b><\/td>\n<td>100%<\/td>\n<\/tr>\n<\/tbody>\n<\/table><\/div>\n<p>  <!-- ===== 20230605 ===== --><\/p>\n<div class=\"hx-section\">\n<h3><span class=\"badge\">20230605<\/span>Actin-Alexa532 \u6d53\u5ea6\u4f18\u5316 \u00b7 \u5173\u952e\u7ed3\u8bba<\/h3>\n<h4>\u539f\u6599\u8bb0\u5f55<\/h4>\n<div class=\"hx-fig-grid cols-2\">\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/WechatIMG652.webp\" alt=\"\u7ec6\u80de\"><figcaption>\u7ec6\u80de\uff08\u56fa\u5b9a \u2192 quench \u2192 \u67d3\u8272\uff09<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/WechatIMG651.webp\" alt=\"\u67d3\u6599 + \u8017\u6750\"><figcaption>\u67d3\u6599\u4e0e\u8017\u6750\uff08-20 \u2103 \u68af\u5ea6\u878d\u5316\uff0c\u5c01\u95ed\u6db2\u73b0\u7528\u73b0\u914d\uff09<\/figcaption><\/figure>\n<\/p><\/div>\n<p><b>\u5b9e\u9a8c\u6761\u4ef6\uff1a<\/b>\u73af\u5883\u6e29\u5ea6 27 \u2103 \u00b7 \u67d3\u8272\u65f6\u95f4 3 h<\/p>\n<table class=\"hx-table\">\n<thead>\n<tr>\n<th>\u6837\u54c1<\/th>\n<th>S01<\/th>\n<th>S02<\/th>\n<th>S03 \u2b50 \u6700\u4f18<\/th>\n<th>S04<\/th>\n<th>S05 \u6b63\u5bf9\u7167<\/th>\n<\/tr>\n<\/thead>\n<tbody>\n<tr>\n<td><b>\u67d3\u6599\u6d53\u5ea6<\/b><\/td>\n<td>50%<\/td>\n<td>100%<\/td>\n<td>200%<\/td>\n<td>300%<\/td>\n<td>488 \u6807\u51c6<\/td>\n<\/tr>\n<tr>\n<td><b>\u914d\u7f6e\u4f53\u7cfb<\/b><\/td>\n<td>\u67d3\u6599\uff08\u7532\u9187\uff0912.5 \u03bcL<br \/>+ Buffer 12.5 \u03bcL<\/td>\n<td>\u67d3\u6599\uff08DMSO\uff092.5 \u03bcL<br \/>+ Buffer 22.5 \u03bcL<\/td>\n<td>\u67d3\u6599\uff08DMSO\uff095 \u03bcL<br \/>+ Buffer 20 \u03bcL<\/td>\n<td>\u67d3\u6599\uff08DMSO\uff097.5 \u03bcL<br \/>+ Buffer 17.5 \u03bcL<\/td>\n<td>488 1 \u03bcL<br \/>+ Buffer 24 \u03bcL<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<h4>20\u00d7 \u65f6\u5e8f cycle<\/h4>\n<div class=\"hx-fig-grid cols-2\">\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230606-S02-cycle-CS.webp\" alt=\"S03 cycle\"><figcaption>S03\uff08200%\uff09\u00b7 20\u00d7 cycle<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230606-S01-cycle-CS.webp\" alt=\"S04 cycle\"><figcaption>S04\uff08300%\uff09\u00b7 20\u00d7 cycle<\/figcaption><\/figure>\n<\/p><\/div>\n<h4>60\u00d7 \u5171\u805a\u7126\u5355\u5e27\uff086.5 \u65e5\uff09<\/h4>\n<div class=\"hx-fig-grid cols-4\">\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230605-S01-C01-CS.webp\" alt=\"S01\"><figcaption>S01 \u00b7 50%<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230605-S02-C01-CS.webp\" alt=\"S02\"><figcaption>S02 \u00b7 100%<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230605-S03-C01-CS.webp\" alt=\"S03\"><figcaption>S03 \u00b7 200% \u2b50<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230605-S04-C01-CS.webp\" alt=\"S04\"><figcaption>S04 \u00b7 300%<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230605-S05-C01-488-CS.webp\" alt=\"S05\"><figcaption>S05 \u00b7 488 \u6b63\u5bf9\u7167<\/figcaption><\/figure>\n<\/p><\/div>\n<h4>60\u00d7 532 \u590d\u6d4b\uff086.6 \u65e5\uff0cS03\/S04\uff09<\/h4>\n<div class=\"hx-fig-grid cols-3\">\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230606-S02-C01-532-CS.webp\" alt=\"S03 532\"><figcaption>S03 \u00b7 200% \u00b7 532<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230606-S02-C02-532-CS.webp\" alt=\"S03 532 C2\"><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230606-S02-C03-532-CS.webp\" alt=\"S03 532 C3\"><\/figure>\n<\/p><\/div>\n<div class=\"hx-callout\">\n      \u2b50 <b>\u5173\u952e\u7ed3\u8bba\uff1a<\/b>\u5f53\u6d53\u5ea6\u8fbe\u5230 <b>200%<\/b> \u65f6\u67d3\u8272\u6548\u679c\u6700\u4f18\uff08\u7070\u5ea6\u5cf0\u503c 4095\uff09\uff0c300% \u65f6\u80cc\u666f\u8367\u5149\u8fc7\u91cd\u3002<br \/>\n      \u4ece\u67d3\u6599\u81ea\u8eab\u770b\uff0c<b>\u8be5\u6279\u6b21 Alexa 532 \u67d3\u6599\u8d28\u91cf\u8f83\u5dee<\/b>\uff0c\u9700 <b>50\u00d7<\/b> \u4e8e\u6b63\u5bf9\u7167\u67d3\u6599\u6d53\u5ea6\u624d\u80fd\u8fbe\u5230\u76f8\u540c\u6548\u679c\uff0c\u5efa\u8bae\u66f4\u6362\u67d3\u6599\u3002\n    <\/div>\n<\/p><\/div>\n<p>  <!-- ===== 20230614 ===== --><\/p>\n<div class=\"hx-section\">\n<h3><span class=\"badge\">20230614<\/span>Actin-Alexa532 \u6837\u54c1\u68c0\u6d4b\u62a5\u544a<\/h3>\n<h4>\u6807\u8bb0\u539f\u6599<\/h4>\n<div class=\"hx-fig-grid cols-2\">\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/WechatIMG729.webp\" alt=\"\u67d3\u6599 1\"><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/WechatIMG728.webp\" alt=\"\u67d3\u6599 2\"><\/figure>\n<\/p><\/div>\n<h4>\u6210\u50cf\u7ed3\u679c\uff08S1 \u00b7 60\u00d7 532 + \u660e\u573a BF\uff09<\/h4>\n<div class=\"hx-fig-grid cols-4\">\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230620-S01_01.webp\" alt=\"C1 532\"><figcaption>C1 \u00b7 532<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230620-S01_C2.webp\" alt=\"C2 532\"><figcaption>C2 \u00b7 532<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230620-S01_C3.webp\" alt=\"C3 532\"><figcaption>C3 \u00b7 532<\/figcaption><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230620-S01_C4.webp\" alt=\"C4 532\"><figcaption>C4 \u00b7 532<\/figcaption><\/figure>\n<\/p><\/div>\n<\/p><\/div>\n<p>  <!-- ===== 20230809 ===== --><\/p>\n<div class=\"hx-section\">\n<h3><span class=\"badge\">20230809<\/span>Actin-Alexa532 \u6837\u54c1\u68c0\u6d4b\u8be6\u60c5<\/h3>\n<div class=\"hx-fig-grid cols-2\">\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/Untitled.webp\" alt=\"20x \u6982\u89c8\"><figcaption>20230809-S01 \u00b7 20\u00d7 \u6982\u89c8<\/figcaption><\/figure>\n<div><\/div>\n<\/p><\/div>\n<h4>60\u00d7 532 \u6210\u50cf<\/h4>\n<div class=\"hx-fig-grid cols-3\">\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230809-S01-C01-561.webp\" alt=\"C01 561\"><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230809-S01-C02-561.webp\" alt=\"C02 561\"><\/figure>\n<figure class=\"hx-figure\"><img decoding=\"async\" src=\"https:\/\/www.hxicollege.com\/wp-content\/uploads\/2026\/07\/1641\/20230809-S01-C03-561.webp\" alt=\"C03 561\"><\/figure>\n<\/p><\/div>\n<\/p><\/div>\n<p>  <!-- ===== \u7ec6\u80de\u57f9\u517b + \u67d3\u8272 Protocol ===== --><\/p>\n<div class=\"hx-section\">\n<h3><span class=\"badge\">Protocol<\/span>\u5b9e\u9a8c\u6d41\u7a0b<\/h3>\n<div class=\"hx-protocol\">\n<p class=\"p-title\"><span class=\"ico\">\ud83e\uddea<\/span><span>\u7ec6\u80de\u722c\u7247\u8868\u9762\u5904\u7406<\/span><\/p>\n<ol>\n<li>\u5c06\u6d01\u51c0\u7684\u7ec6\u80de\u722c\u7247\u653e\u5165\u516d\u5b54\u677f\u6216 3.5 cm \u57f9\u517b\u76bf\u4e2d\uff0c\u52a0\u5165 1\u20132 mL \u5b8c\u5168\u57f9\u517b\u57fa\u4f7f\u4e4b\u6d78\u6da6\u722c\u7247\uff1b<\/li>\n<li>\u653e\u5165\u57f9\u517b\u7bb1\u5b75\u80b2 <b>12 h<\/b> \u4ee5\u4e0a\uff0c\u4f7f\u7528\u524d\u5438\u9664\u57f9\u517b\u57fa\u3002<\/li>\n<\/ol><\/div>\n<div class=\"hx-protocol\">\n<p class=\"p-title\"><span class=\"ico\">\ud83e\uddeb<\/span><span>\u6210\u50cf\u7ec6\u80de\u63a5\u79cd<\/span><\/p>\n<ol>\n<li>\u53d6\u7ec6\u80de\u72b6\u6001\u6700\u4f18\u7684\u57f9\u517b\u76bf\u6d88\u5316\u5904\u7406\uff0c\u6d88\u5316\u79bb\u5fc3\u540e\u7ec6\u80de\u8ba1\u6570\uff0c\u786e\u4fdd\u6bcf\u5b54\uff08\u516d\u5b54\u677f\uff09\/ \u6bcf\u76bf \u5185\u6709 <b>4 \u00d7 10\u2074<\/b> \u4e2a\u6d3b\u7ec6\u80de\uff0c\u8ba1\u7b97\u6240\u9700\u7ec6\u80de\u603b\u91cf\uff1b<\/li>\n<li>\u53d6\u51fa\u6240\u9700\u7ec6\u80de\uff0c\u6309\u6bcf\u5b54\/\u6bcf\u76bf\u52a0 2 mL \u57f9\u517b\u57fa\u8ba1\u7b97\uff0c\u914d\u5236\u6210\u7ec6\u80de\u60ac\u6db2\u5e76\u52a0\u5165\u5230\u5b54\u677f\u6216\u57f9\u517b\u76bf\u4e2d\uff1b<\/li>\n<li>\u6210\u50cf\u7ec6\u80de\u57f9\u517b\u9700 1\u20132 \u5929\uff0c\u89c2\u6d4b\u5230\u7ec6\u80de\u72b6\u6001\u826f\u597d\u4e14\u5355\u4e2a\u7ec6\u80de\u8212\u5c55\u4e0d\u62e5\u6324\u3001\u9762\u79ef\u5927\u3001\u7ec6\u80de\u900f\u5149\u6027\u5f3a\u900f\u660e\u5ea6\u9ad8\u65f6\uff0c\u53ef\u8fdb\u884c\u540e\u7eed\u6210\u50cf\u6b65\u9aa4\u3002<\/li>\n<\/ol><\/div>\n<div class=\"hx-protocol\">\n<p class=\"p-title\"><span class=\"ico\">\ud83c\udfa8<\/span><span>\u67d3\u8272\u6b65\u9aa4<\/span><\/p>\n<ol>\n<li><b>\u7ec6\u80de\u56fa\u5b9a\uff1a<\/b>\u4f7f\u7528 4% PFA \u56fa\u5b9a\u7ec6\u80de\u5f62\u6001\uff1b<\/li>\n<li><b>\u80cc\u666f\u8367\u5149\u6dec\u706d\uff1a<\/b>\u4f7f\u7528\u787c\u6c22\u5316\u94a0\u6d88\u9664\u80cc\u666f\u8367\u5149\uff1b<\/li>\n<li><b>\u7ec6\u80de\u5c01\u95ed\uff1a<\/b>\u4f7f\u7528 BSA \u5904\u7406\u7ec6\u80de\uff0c\u4f7f\u5176\u7ed3\u5408\u4f4d\u70b9\u66f4\u6613\u7ed3\u5408\u67d3\u6599\uff1b<\/li>\n<li><b>\u7ec6\u80de\u67d3\u8272\uff1a<\/b>\u4f7f\u7528 Alexa 532 \u67d3\u6599\u67d3\u8272\u7ec6\u80de\uff0c\u8fc7\u591c\u5b75\u80b2\uff1b<\/li>\n<li><b>\u7ec6\u80de\u540e\u56fa\u5b9a\uff1a<\/b>\u518d\u6b21\u4f7f\u7528 PFA \u56fa\u5b9a\u7ec6\u80de\u5f62\u6001\uff0c\u4f7f\u6837\u7247\u5f62\u6001\u66f4\u52a0\u7a33\u5b9a\u3002<\/li>\n<\/ol>\n<p style=\"margin:10px 0 0;color:#64748b;font-size:13.5px;\">\u26a0\ufe0f \u6ce8\u610f\uff1a\u6bcf\u4e00\u6b65\u7ec6\u80de\u5904\u7406\u540e\u90fd\u9700\u8981\u5bf9\u7ec6\u80de\u8fdb\u884c\u6e05\u6d17\uff0c\u53bb\u9664\u4e0a\u4e00\u6b65\u4f7f\u7528\u7684\u8bd5\u5242\u3002<\/p>\n<\/p><\/div>\n<\/p><\/div>\n<p>  <!-- ===== \u67d3\u8272\u8fc7\u7a0b\u63cf\u8ff0 ===== --><\/p>\n<div class=\"hx-section\">\n<h3><span class=\"badge\">Methods<\/span>\u67d3\u8272\u8fc7\u7a0b\u63cf\u8ff0<\/h3>\n<div class=\"hx-protocol\">\n<p class=\"p-title\"><span class=\"ico\">\ud83e\uddec<\/span><span>Actin staining\uff08\u808c\u52a8\u86cb\u767d\u67d3\u8272\uff09<\/span><\/p>\n<p><b>Cell culture<\/b><\/p>\n<p>The Biologics Standards-Cercopithecus-1 (BSC-1) cell line and NIH\/3T3 cell line were purchased from Pricella Life Technology Co. LTD. BSC-1 cells and NIH\/3T3 cells were cultured in DMEM (Invitrogen, #11965-118) supplemented with 10% fetal bovine serum (FBS) (Gibco, #16010-159). To prevent bacterial contamination, 100 \u03bcg\/mL penicillin and streptomycin (Invitrogen, #15140122) were added in the DMEM medium. Cells were grown under standard cell culture conditions (5% CO\u2082, humidified atmosphere at 37 \u2103). BSC-1 cells were plated on #1.5 glass-bottom dish over 48 h before sample preparation. For cell passage, cells were washed with pre-warmed PBS (Life Technologies, #14190500BT) 3 times and digested with 25% trypsin (Gibco, #25200-056) for 30 s. BSC-1 cell line were tested for potential mycoplasma contamination (MycoAlert, Lonza), and all tests showed negative results.<\/p>\n<p><b>Cell staining<\/b><\/p>\n<p>Cells were grown on 35 mm, #1.5 glass coverslips (Standard Imaging (Beijing) Biotech, STGBD-035-1). To increase cell adhesion, we pre-treated glass-bottom dishes with fibronectin (Invitrogen, #33016015) for 1 h at 37 \u2103. On the day of sample preparation, the cell density should be about 50%\u201370%. Cells were fixed with 37 \u2103 pre-warmed fixation buffer for 10 min, containing 4% paraformaldehyde (EMS) and 0.1% glutaraldehyde in PBS. Then the sample were washed three times with PBS. For quenching the background fluorescence, we incubate the cells with 2 mL 0.1% NaBH\u2084 solution in PBS for 7 min, optionally shake on the shaker (&lt;1 Hz). The sample were washed three times with 2 mL PBS and then incubated for 30 min in PBS containing 5% BSA (Jackson, #001-000-162) and 0.5% Triton X-100 (Fisher Scientific) at 37 \u2103. All dyes were diluted in the 5% BSA + 0.5% triton solution described above. Next, we incubate the sample for 40 min with the appropriate dilution of dyes: phalloidin-Alexa532 (ThermoFisher, A22282) or Acti-Stain 555 Phalloidin (Cytoskeleton, PHDH1-A) at 25 \u2103. After washed 3 times with PBS, cells were fixed with post-fixation buffer for 10 min. The samples were stored in 4 \u2103 in PBS no more than one week.<\/p>\n<\/p><\/div>\n<div class=\"hx-protocol\">\n<p class=\"p-title\"><span class=\"ico\">\ud83e\uddec<\/span><span>Tubulin staining\uff08\u5fae\u7ba1\u67d3\u8272\uff09<\/span><\/p>\n<p><b>Cell culture<\/b><\/p>\n<p>The Biologics Standards-Cercopithecus-1 (BSC-1) cell line was purchased from Pricella Life Technology Co. LTD. BSC-1 cells were cultured in DMEM (Invitrogen, #11965-118) supplemented with 10% fetal bovine serum (FBS) (Gibco, #16010-159). To prevent bacterial contamination, 100 \u03bcg\/mL penicillin and streptomycin (Invitrogen, #15140122) were added in the DMEM medium. Cells were grown under standard cell culture conditions (5% CO\u2082, humidified atmosphere at 37 \u2103). BSC-1 cells were plated on #1.5 glass-bottom dish over 48 h before sample preparation. For cell passage, cells were washed with pre-warmed PBS (Life Technologies, #14190500BT) 3 times and digested with 25% trypsin (Gibco, #25200-056) for 30 s. BSC-1 cell line were tested for potential mycoplasma contamination (MycoAlert, Lonza), and all tests showed negative results.<\/p>\n<p><b>Immunofluorescence staining<\/b><\/p>\n<p>Cells were grown on 35 mm, #1.5 glass coverslips (Standard Imaging (Beijing) Biotech, STGBD-035-1). To increase cell adhesion, we pre-treated glass-bottom dishes with fibronectin (Invitrogen, #33016015) for 1 h at 37 \u2103. On the day of sample preparation, the cell density should be about 50%\u201370%. Cells were fixed with 37 \u2103 pre-warmed fixation buffer for 10 min, containing 4% paraformaldehyde (EMS) and 0.1% glutaraldehyde in PBS. Then the sample were washed three times with PBS. For quenching the background fluorescence, we incubate the cells with 2 mL 0.1% NaBH\u2084 solution in PBS for 7 min, optionally shake on the shaker (&lt;1 Hz). The sample were washed three times with 2 mL PBS and then incubated for 30 min in PBS containing 5% BSA (Jackson, #001-000-162) and 0.5% Triton X-100 (Fisher Scientific) at 37 \u2103. All antibodies were diluted in the 5% BSA + 0.5% triton solution described above. Next, we incubate the sample for 12 hours the appropriate dilution of primary antibodies: beta-tubulin (DSHB-E7) at 4 \u2103. After primary antibodies incubation, the cells were washed 5 min with 2 mL PBS for three times. Secondary antibodies were incubated for 60 min with the appropriate dilutions of secondary antibodies (Cy3b secondary antibodies were conjugated in our laboratory) at 25 \u2103, the sample were protected from light. After washed 3 times with PBS, cells were fixed with post-fixation buffer for 10 min. The samples were stored in 4 \u2103 in PBS no more than one week.<\/p>\n<\/p><\/div>\n<\/p><\/div>\n<p>  <!-- ===== \u4e0b\u4e00\u6b65\u884c\u52a8 ===== --><\/p>\n<div class=\"hx-section\">\n<h3><span class=\"badge\">Next<\/span>\u4e0b\u4e00\u6b65\u884c\u52a8<\/h3>\n<p>\u2705 <b>\u5df2\u5b8c\u6210\uff1a<\/b>\u5728\u57fa\u6069\u58eb\u4e0a\u5efa\u7acb\u6837\u54c1\u4eae\u5ea6\u7684 positive control\uff0c\u4f5c\u4e3a\u540e\u7eed\u6279\u6b21\u7684\u5bf9\u7167\u57fa\u51c6\u3002<\/p>\n<table class=\"hx-table\">\n<thead>\n<tr>\n<th>\u6d53\u5ea6\uff08\u7532\u9187\uff09<\/th>\n<th>20%<\/th>\n<th>50%<\/th>\n<th>75%<\/th>\n<th>100%<\/th>\n<th>DMSO \u6eb6<\/th>\n<th>150%<\/th>\n<th>200%<\/th>\n<th>250%<\/th>\n<th>300%<\/th>\n<\/tr>\n<\/thead>\n<tbody>\n<tr>\n<td><b>\u5bf9\u5e94 \u03bcM<\/b><\/td>\n<td>6.6 \u03bcM<\/td>\n<td colspan=\"4\"><\/td>\n<td>66 \u03bcM<\/td>\n<td colspan=\"3\"><\/td>\n<\/tr>\n<tr>\n<td><b>\u662f\u5426\u9700\u8981<\/b><\/td>\n<td>\u4e0d\u9700\u8981<\/td>\n<td>1:1<\/td>\n<td>\u4e0d\u9700\u8981<\/td>\n<td>\u6539\u4e3a DMSO Stock<\/td>\n<td>\u2014<\/td>\n<td>\u4e0d\u9700\u8981<\/td>\n<td>\u4e0d\u9700\u8981<\/td>\n<td>\u4e0d\u9700\u8981<\/td>\n<td>\u2014<\/td>\n<\/tr>\n<tr>\n<td><b>phalloidin Alexa532 \u7528\u91cf<\/b><\/td>\n<td>5 \u03bcL<\/td>\n<td>12.5 \u03bcL<\/td>\n<td>20 \u03bcL<\/td>\n<td>25 \u03bcL<\/td>\n<td>\u2014<\/td>\n<td>3.75 \u03bcL<\/td>\n<td>5 \u03bcL<\/td>\n<td>6.25 \u03bcL<\/td>\n<td>7.5 \u03bcL<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p><b>\u7528\u91cf\u6c47\u603b\uff1a<\/b>\u7532\u9187\u914d\u7f6e\u65b9\u6848\u5171\u9700 57.5 \u03bcL\uff1bDMSO \u6d53\u7f29\u65b9\u6848\u5171\u9700 22.5 \u03bcL\u3002<\/p>\n<p><b>\u5efa\u8bae\uff1a<\/b>\u5c06\u7532\u9187\u914d\u7f6e\u7684\u67d3\u6599 225 \u03bcL \u6d53\u7f29\u81f3 DMSO \u914d\u7f6e\u7684 22.5 \u03bcL\uff0c\u5efa\u8bae\u4e00\u6b21\u6027\u914d\u7f6e 200 \u03bcL\uff08\u2192 \u5b9e\u9645\u53ef\u7528 20 \u03bcL\uff09\u3002<\/p>\n<\/p><\/div>\n<p class=\"hx-end\">\u2014\u2014 \u6848\u4f8b 03 \u00b7 \u5ba2\u6237\u6848\u4f8b \u00b7 \u534e\u4e1c\u5e08\u8303\u5927\u5b66-\u9ec4\u6b63\u9f50\u8001\u5e08\u6837\u54c1\u68c0\u6d4b\u62a5\u544a \u2014\u2014<\/p>\n<\/div>\n","protected":false},"excerpt":{"rendered":"<p>\u9879\u76ee\u8be6\u60c5\uff1a<br \/>\n\u534e\u4e1c\u5e08\u8303\u5927\u5b66-\u9ec4\u6b63\u9f50\u8001\u5e08\u5b9a\u5236\u9879\u76ee<\/p>\n<p>\u6807\u8bb0\u539f\u6599\u8bb0\u5f55<br \/>\nActi-Stain 555 Phalloidin<br \/>\nhttps:\/\/www.cytoskeleton.com\/phdh1<\/p>\n<p>Alexa Fluor\u2122 532\u9b3c\u7b14\u73af\u80bd<br \/>\n\u6fc0\u53d1\/\u53d1\u5c04\u6ce2\u957f\uff1a531\/554 nm<br \/>\n[\u56fe\u7247]<br \/>\nhttps:\/\/www.thermofisher.cn\/order\/catalog\/product\/A22282<br \/>\nIn DMSO\uff0c150ul 66um stock<br \/>\nIn methano\uff0c1.5ml  6.6um stock<br \/>\n\u9009\u62e9\u7528methano\u6eb6\u89e3\uff0c\u9700\u8981\u505a\u6d53\u5ea6titration<br \/>\nMAN0001777_Phalloidins_UG.pdf<\/p>\n<p>\u68c0\u6d4b\u8bbe\u5907Olympus FV3000<\/p>\n","protected":false},"author":1,"featured_media":1649,"comment_status":"open","ping_status":"open","sticky":true,"template":"","format":"standard","meta":{"footnotes":""},"categories":[52,55],"tags":[48],"class_list":["post-1641","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-customer-case","category-select-case","tag-main"],"_links":{"self":[{"href":"https:\/\/www.hxicollege.com\/index.php\/wp-json\/wp\/v2\/posts\/1641","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.hxicollege.com\/index.php\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.hxicollege.com\/index.php\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.hxicollege.com\/index.php\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.hxicollege.com\/index.php\/wp-json\/wp\/v2\/comments?post=1641"}],"version-history":[{"count":2,"href":"https:\/\/www.hxicollege.com\/index.php\/wp-json\/wp\/v2\/posts\/1641\/revisions"}],"predecessor-version":[{"id":1650,"href":"https:\/\/www.hxicollege.com\/index.php\/wp-json\/wp\/v2\/posts\/1641\/revisions\/1650"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.hxicollege.com\/index.php\/wp-json\/wp\/v2\/media\/1649"}],"wp:attachment":[{"href":"https:\/\/www.hxicollege.com\/index.php\/wp-json\/wp\/v2\/media?parent=1641"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.hxicollege.com\/index.php\/wp-json\/wp\/v2\/categories?post=1641"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.hxicollege.com\/index.php\/wp-json\/wp\/v2\/tags?post=1641"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}