\n\u7f16\u53f7<\/span><\/td>\n | \u7ec4\u5206<\/span><\/td>\n | \u4fdd\u5b58\u65b9\u6cd5<\/span><\/td>\n | \n \u4ea7\u54c1\u7f16\u53f7\/\u89c4\u683c<\/span><\/p>\n<\/td>\n<\/tr>\n\nMX3012-500T<\/span><\/td>\n | MX3012-1000T<\/span><\/td>\n<\/tr>\n\nMX3012-A<\/span><\/td>\n | Calcein AM Solution(4 mM)<\/span><\/td>\n | -20\u00baC\u907f\u5149<\/span><\/td>\n | 50\u03bcl<\/span><\/td>\n | 50\u03bcl\u00d72<\/span><\/td>\n<\/tr>\n\nMX3012-B<\/span><\/td>\n | PI Solution\uff081.5 mM\uff09<\/span><\/td>\n | -20\u00baC\u907f\u5149<\/span><\/td>\n | 200\u03bcl<\/span><\/td>\n | 200\u03bcl\u00d72<\/span><\/td>\n<\/tr>\n\nMX3012-C<\/span><\/td>\n | 10\u00d7Assay Buffer<\/span><\/td>\n | -20\u00baC~4\u2103<\/span><\/td>\n | 50ml<\/span><\/td>\n | 100ml<\/span><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n \u4fdd\u5b58\u4e0e\u8fd0\u8f93\u65b9\u6cd5<\/strong><\/span><\/p>\n\u4fdd\u5b58\uff1a-20\u2103\u907f\u5149\u4fdd\u5b58\uff0c\u6709\u6548\u671f\u4e00\u5e74\u3002<\/span><\/p>\n\u8fd0\u8f93\uff1a\u51b0\u888b\u8fd0\u8f93\u3002<\/span><\/p>\n\u6ce8\u610f\u4e8b\u9879<\/strong><\/span><\/p>\n1\uff09\u7531\u4e8eCalcein-AM\u5bf9\u6e7f\u5ea6\u975e\u5e38\u654f\u611f\uff0c\u82e5\u662fCalcein-AM\u6eb6\u6db2\u6bcf\u6b21\u53d6\u5b8c\u9700\u8981\u91cf\u540e\uff0c\u5fc5\u987b\u7d27\u7d27\u5bc6\u5c01\u76d6\u5b50\u3002\u5efa\u8bae\u6839\u636e\u5355\u6b21\u7528\u91cf\uff0c\u5206\u88c5\u5bc6\u5c01\u4fdd\u5b58\u3002Calcein-AM\u5de5\u4f5c\u6db2\u5fc5\u987b\u73b0\u914d\u73b0\u7528\u3002<\/span><\/p>\n2\uff09\u7898\u5316\u4e19\u5576\uff08PI\uff09\u6709\u4e00\u5b9a\u7684\u81f4\u764c\u6027\uff0c\u64cd\u4f5c\u65f6\u4e00\u5b9a\u8981\u6ce8\u610f\u9632\u62a4\u3002\u82e5\u63a5\u89e6\u5230\u76ae\u80a4\uff0c\u9700\u8981\u7acb\u5373\u7528\u81ea\u6765\u6c34\u6e05\u6d17\u3002<\/span><\/p>\n3\uff09\u4e3a\u4e86\u60a8\u7684\u5b89\u5168\u548c\u5065\u5eb7\uff0c\u8bf7\u7a7f\u5b9e\u9a8c\u670d\u5e76\u6234\u4e00\u6b21\u6027\u624b\u5957\u64cd\u4f5c\u3002<\/span><\/p>\n\u4f7f\u7528\u65b9\u6cd5<\/strong><\/span><\/p>\n\u4e00\u3001\u8bd5\u5242\u51c6\u5907<\/span><\/p>\n1.1 1\u00d7\u53cd\u5e94\u7f13\u51b2\u6db2\uff08Assay Buffer\uff09\u7684\u914d\u5236<\/span><\/p>\n\u4ece\u4f4e\u6e29\u51b0\u7bb1\u5185\u53d6\u51fa10\u00d7Assay Buffer\uff0c\u6839\u636e\u5355\u6b21\u7528\u91cf\u65e0\u83cc\u6761\u4ef6\u53d6\u51fa\u9002\u91cf\uff0c\u7528\u53bb\u79bb\u5b50\u6c34\uff08dH2O\uff09\u505a10\u500d\u7a00\u91ca\u4ee5\u5f97\u52301\u00d7Assay Buffer\u3002<\/span><\/p>\n1.2 1\u00d7\u67d3\u8272\u5de5\u4f5c\u6db2\uff08Stain Buffer\uff09\u7684\u914d\u5236<\/span><\/p>\n1\uff09 \u5148\u5c06\u4f4e\u6e29\u4fdd\u5b58\u7684Calcein AM\u6eb6\u6db2 (4 mM)\u548cPI\u6eb6\u6db2\uff081.5 mM\uff09\u5ba4\u6e29\u56de\u6e29\u81f3\u5c1120min\uff0c\u4f7f\u5176\u5145\u5206\u878d\u5316\u3002\u3010\u6ce8\u610f\uff1a\u7b2c\u4e00\u6b21\u4f7f\u7528\u65f6\u5efa\u8bae\u5bf9\u50a8\u5b58\u6db2\u6839\u636e\u5355\u6b21\u7528\u91cf\u5206\u88c5\uff0c\u7279\u522b\u662fCalcein AM\uff0c\u4ee5\u51cf\u5c11\u53cd\u590d\u51bb\u878d\u6b21\u6570\u3002\u3011<\/span><\/p>\n2\uff09 \u53d65\u00b5l Calcein AM\u6eb6\u6db2 (4 mM)\u548c15\u00b5l PI\u6eb6\u6db2\uff081.5 mM\uff09\u52a0\u51655ml 1\u00d7Assay Buffer\uff0c\u5145\u5206\u6df7\u5300\u3002\u6b64\u65f6\u5f97\u5230Calcein AM\u7684\u5de5\u4f5c\u6db2\u6d53\u5ea6\u4e3a4\u03bcM\uff0cPI\u7684\u5de5\u4f5c\u6db2\u6d53\u5ea6\u4e3a4.5\u03bcM\u3002\u7531\u4e8e\u4e0d\u540c\u7ec6\u80de\u7cfb\u7684\u6700\u4f73\u67d3\u8272\u6761\u4ef6\u4e0d\u540c\uff0c\u521d\u6b21\u5b9e\u9a8c\u5efa\u8bae\u505a\u68af\u5ea6\u5b9e\u9a8c\uff0c\u4ee5\u786e\u5b9a Calcein-AM\u548cPI\u7684\u6700\u9002\u6d53\u5ea6\u3002\u68af\u5ea6\u7b5b\u9009\u7684\u539f\u5219\u4e3a\u4f7f\u7528\u6700\u4f4e\u7684\u63a2\u9488\u6d53\u5ea6\u5f97\u5230\u6700\u597d\u7684\u8367\u5149\u7ed3\u679c\u3002\u3010\u6ce8\u610f\uff1a\u67d3\u8272\u5de5\u4f5c\u6db2\u5fc5\u987b\u73b0\u914d\u73b0\u7528\uff0c\u5e76\u4e14\u5728\u5f53\u5929\u7528\u5b8c\u3002\u3011<\/span><\/p>\n\u4e8c\u3001\u67d3\u8272\u6b65\u9aa4<\/span><\/p>\n2.1 \u5bf9\u4e8e\u8d34\u58c1\u7ec6\u80de\uff0c\u5148\u7528\u7ec6\u80de\u522e\u5200\u6216\u8005\u80f0\u9176-EDTA\u6d88\u5316\u7ec6\u80de\uff0c\u4e4b\u540e\u79bb\u5fc3\u6536\u96c6\u7ec6\u80de\uff081000 rpm\uff0c3min\uff09\u3002<\/span><\/p>\n\u5bf9\u4e8e\u60ac\u6d6e\u7ec6\u80de\uff0c\u76f4\u63a5\u79bb\u5fc3\uff081000 rpm\uff0c3min\uff09\u6536\u96c6\u7ec6\u80de\u3002<\/span><\/p>\n2.2 \u53bb\u4e0a\u6e05\uff0c\u75281\u00d7Assay Buffer\u5145\u5206\u6e05\u6d17\u7ec6\u80de2~3\u6b21\uff0c\u4ee5\u5145\u5206\u53bb\u9664\u6b8b\u7559\u7684\u916f\u9176\u6d3b\u6027\u3002<\/span><\/p>\n2.3 \u75281\u00d7Assay Buffer\u5236\u5907\u7ec6\u80de\u60ac\u6db2\uff0c\u4f7f\u5176\u5bc6\u5ea6\u4e3a1\u00d7105~1\u00d7106\u7ec6\u80de\/ml\u3002<\/span><\/p>\n2.4 \u53d6100\u00b5l\u67d3\u8272\u5de5\u4f5c\u6db2\u52a0\u5165200\u00b5l\u7ec6\u80de\u60ac\u6db2\u5185\uff0c\u6df7\u5300\uff0c37\u2103\u5b75\u80b215min\u3002<\/span><\/p>\n\u3010\u6ce8\u610f\uff1a\u5982\u679c\u9700\u8981\uff0c\u53ef\u5ef6\u957f\u5b75\u80b2\u65f6\u95f4\u81f330min\u3002\u3011<\/span><\/p>\n2.5 \u8367\u5149\u663e\u5fae\u955c\u4e0b\u4f7f\u7528490\u00b110 nm\u6fc0\u53d1\u6ee4\u7247\u540c\u65f6\u68c0\u6d4b\u6d3b\u7ec6\u80de\uff08\u9ec4\u7eff\u8272\u8367\u5149\uff09\u4ee5\u53ca\u6b7b\u7ec6\u80de\uff08\u7ea2\u8272\u8367\u5149\uff09\u3002\u53e6\u5916\uff0c\u4f7f\u7528545nm\u7684\u53d1\u5c04\u6ee4\u7247\u4ec5\u80fd\u89c2\u5bdf\u5230\u6b7b\u7ec6\u80de\u3002\u4e5f\u53ef\u4ee5\u76f4\u63a5\u5728\u8367\u5149\u9176\u6807\u4eea\u4e0b\u4f7f\u7528\u5408\u9002\u7684\u6ee4\u7247\u8fdb\u884c\u68c0\u6d4b\u3002<\/span><\/p>\n\u3010\u6ce8\u610f\u3011\uff1a\u53ef\u4ee5\u4f7f\u7528\u4ee5\u4e0b\u65b9\u6cd5\u6765\u4f18\u5316\u5f97\u5230\u4e24\u79cd\u8367\u5149\u67d3\u6599\u7684\u6700\u4f73\u5de5\u4f5c\u6d53\u5ea6\u3002<\/span><\/p>\na) \u75280.1%\u7682\u7d20\u6216\u80050.1-0.5%\u5730\u9ad8\u8f9b\u5b75\u80b2\u7ec6\u80de10min\uff0c\u6216\u8005\u752870%\u4e59\u9187\u5b75\u80b2\u7ec6\u80de30min\uff0c\u4ece\u800c\u5236\u5907\u6b7b\u7ec6\u80de\uff1b<\/span><\/p>\nb) \u75280.1-10\u00b5M\u7684PI\u6eb6\u6db2\u8fdb\u884c\u6b7b\u7ec6\u80de\u67d3\u8272\uff0c\u4ee5\u5f97\u5230\u4ec5\u4ec5\u5bf9\u7ec6\u80de\u6838\u67d3\u8272\uff0c\u800c\u4e0d\u4f1a\u5bf9\u7ec6\u80de\u8d28\u67d3\u8272\u7684\u6700\u4f73\u5de5\u4f5c\u6d53\u5ea6\u3002<\/span><\/p>\nc) \u75280.1-10\u00b5M\u7684Calcein AM\u8fdb\u884c\u6b7b\u7ec6\u80de\u67d3\u8272\uff0c\u4ee5\u5f97\u5230\u4e0d\u4f1a\u5bf9\u7ec6\u80de\u8d28\u67d3\u8272\u7684\u6700\u4f73\u5de5\u4f5c\u6d53\u5ea6\u3002\u7136\u540e\u7528\u6b64\u6d53\u5ea6\u8fdb\u884c\u6d3b\u7ec6\u80de\u67d3\u8272\uff0c\u53bb\u89c2\u5bdf\u662f\u5426\u6d3b\u7ec6\u80de\u80fd\u88ab\u67d3\u8272\u3002<\/span><\/p>\n\u6d4b\u5b9a\u65b9\u6cd5\u4e0e\u7ed3\u679c\u5448\u73b0 <\/strong><\/span><\/p>\n1\uff09 \u8367\u5149\u663e\u5fae\u955c\uff08Fluorescent Microscopy\uff09 <\/strong><\/span><\/p>\n\u68c0\u6d4b\u65b9\u6cd5\uff1a490\u00b110 nm\u6fc0\u53d1\u80fd\u540c\u65f6\u770b\u5230\u5448\u9ec4-\u7eff\u8272\u8367\u5149\u7684\u6d3b\u7ec6\u80de\u548c\u7ea2\u8272\u7684\u6b7b\u7ec6\u80de\u3002\u53e6\uff0c545nm\u6fc0\u53d1\u53ef\u80fd\u53ea\u80fd\u770b\u5230\u5448\u7ea2\u8272\u7684\u6b7b\u7ec6\u80de\u3002<\/span><\/p>\n <\/strong><\/span><\/p>\nFig 1. Fluorescence microscopy images of calcein-AM (green) and propidium iodide (red). The macrophages differentiated into osteoclasts on (a) nt-TiO2 and (b) nt-TiO2-P for 4 days. Cells were then examined by fluorescence microscopy (Axioplan 2, Carl Zeiss, Oberkochen, Germany) with 490-nm excitation for the simultaneous monitoring of viable and dead cells. [Reference: Tae-Hyung Koo et al. Immobilization of pamidronic acids on the nanotube surface of titanium discs and their interaction with bone cells. Nanoscale Res Lett. 2013; 8(1): 124.]<\/span><\/p>\n2\uff09 \u8367\u5149\u9176\u6807\u4eea\uff08Fluorescent Plate Reader\uff09 <\/strong><\/span><\/p>\n\u68c0\u6d4b\u65b9\u6cd5\uff1a96\u5b54\u900f\u660e\u5e95\u7684\u9ed1\u8272\u9176\u6807\u677f\uff0896-well black plate with clear bottom\uff09\uff0cCalcein\u68c0\u6d4b\u7684\u6fc0\u53d1\u6ce2\u957f\u4e3a490nm\uff0c\u53d1\u5c04\u6ce2\u957f\u4e3a520nm\uff1bPI\u68c0\u6d4b\u7684\u6fc0\u53d1\u6ce2\u957f\u4e3a530nm\uff0c\u53d1\u5c04\u6ce2\u957f\u4e3a617nm\uff1b<\/span><\/p>\n3\uff09 \u6d41\u5f0f\u7ec6\u80de\u4eea\uff08Flow cytometer\uff09 <\/strong><\/span><\/p>\n\u68c0\u6d4b\u65b9\u6cd5\uff1a\u6d41\u5f0f\u7ec6\u80de\u4eea\uff0cCalcein\u68c0\u6d4b\u7684\u6fc0\u53d1\/\u53d1\u5c04\u6ce2\u957f\u4e3a488\/535nm\uff0cPI\u7684\u6fc0\u53d1\u548c\u53d1\u5c04\u6ce2\u957f\u4e3a488\/620nm\u3002<\/span><\/p>\n <\/strong><\/span><\/p>\nFig 3. Quantification of calcein and PI staining in WT cells and in cells incubated with increasing concentrations of miltefosine: 20\u03bcM, 40\u03bcM or 100\u03bcM for 24h. [Reference: Basmaciyan L et al. Calcein+\/PI- as an early apoptotic feature in Leishmania. PLoS One. 2017 Nov 7;12(11):e0187756.]<\/span><\/p>\n\u5e38\u89c1\u95ee\u9898<\/strong> <\/b><\/span><\/p>\n1\u3001 \u8be5\u8bd5\u5242\u76d2\u9002\u7528\u4efb\u4f55\u7ec6\u80de\u7c7b\u578b\uff1f<\/span><\/p>\n\u57fa\u672c\u4e0a\u9002\u7528\u4e8e\u4efb\u4f55\u542b\u916f\u9176\u6d3b\u6027\u7684\u52a8\u7269\u7ec6\u80de\u3002\u690d\u7269\u548c\u7ec6\u83cc\u7ec6\u80de\u56e0\u542b\u6709\u7ec6\u80de\u58c1\uff0cCalcein-AM\u4e0d\u80fd\u8fdb\u5165\u56e0\u6b64\u65e0\u6cd5\u67d3\u8272\u3002\u6709\u53ef\u80fd\u5bf9\u539f\u751f\u8d28\u4f53\u8fdb\u884c\u67d3\u8272\u3002<\/span><\/p>\n2\u3001 \u7528\u76f8\u540c\u6d53\u5ea6\u6709\u53ef\u80fd\u5bf9\u6240\u6709\u54fa\u4e73\u52a8\u7269\u7ec6\u80de\u8fdb\u884c\u67d3\u8272\uff1f<\/span><\/p>\n\u5bf9\u6240\u6709\u7ec6\u80de\u90fd\u7528\u76f8\u540c\u7684\u6d53\u5ea6\u662f\u5f88\u4e0d\u5408\u7406\u7684\u3002Calcein-AM\u548cPI\u7684\u6700\u4f73\u5de5\u4f5c\u6d53\u5ea6\u6839\u636e\u7ec6\u80de\u7c7b\u578b\u5dee\u5f02\u5f88\u5927\u3002\u6709\u5fc5\u8981\u6839\u636e\u5177\u4f53\u7ec6\u80de\u6765\u786e\u5b9a\u6700\u4f73\u67d3\u6599\u6d53\u5ea6\u3002\u53c2\u8003\u8bf4\u660e\u4e66\u67d3\u6599\u5de5\u4f5c\u6d53\u5ea6\u4f18\u5316\u65b9\u6cd5\u3002<\/span><\/p>\n3\u3001 Calcein-AM\u5bf9\u7ec6\u80de\u6709\u6bd2\uff1f<\/span><\/p>\n\u4e0e\u5176\u4ed6\u76f8\u4f3c\u7684\u6d3b\u7ec6\u80de\u67d3\u6599\u6bd4\u8f83\uff0cCalcein-AM\u57fa\u672c\u65e0\u6bd2\uff08\u6bd2\u6027\u6700\u5c0f\uff09\u3002\u89c1\u6587\u732eEL.S.D.Clerck,et.al., J.Immunol.Methods, 172,115-124(1994)<\/span><\/p>\n\u76f8\u5173\u4ea7\u54c1<\/strong><\/span><\/p>\n\n\n\n\u8d27\u53f7<\/span><\/td>\n | \u540d\u79f0<\/span><\/td>\n | \u89c4\u683c<\/span><\/td>\n<\/tr>\n\nMX3011-50UG<\/span><\/td>\n | Calcein, AM, Ultra Pure Grade \u9499\u9ec4\u7eff\u7d20\uff08\u7eff\u8272\uff09<\/span><\/td>\n | 50\u03bcg<\/span><\/td>\n<\/tr>\n\nMX3012-500T<\/span><\/td>\n | Calcein AM\/PI Double Stain Kit \u6d3b\u7ec6\u80de\/\u6b7b\u7ec6\u80de\u53cc\u67d3\u8bd5\u5242\u76d2<\/span><\/td>\n | 500T<\/span><\/td>\n<\/tr>\n\nMX3013-1MG<\/span><\/td>\n | Calcein Red, AM \u9499\u9ec4\u7eff\u7d20\u7ea2\uff08\u7ea2\u8272\uff09<\/span><\/td>\n | 1mg<\/span><\/td>\n<\/tr>\n\nMX3014-1MG<\/span><\/td>\n | Calcein Deep Red Acetate \u9499\u9ec4\u7eff\u7d20\u6df1\u7ea2\u918b\u9178\u76d0<\/span><\/td>\n | 1mg<\/span><\/td>\n<\/tr>\n\nMZ8471-1G<\/span><\/td>\n | Probenecid \u4e19\u78fa\u8212<\/span><\/td>\n | 1g<\/span><\/td>\n<\/tr>\n\nMZ8472-154MG<\/span><\/td>\n | Probenecid Sodium, Water Soluble \u4e19\u78fa\u8212\u94a0\uff08\u6c34\u6eb6\u6027\uff09<\/span><\/td>\n | 2\u00d777mg<\/span><\/td>\n<\/tr>\n\nMX4205-10MG<\/span><\/td>\n | Propidium Iodide \u7898\u5316\u4e19\u5576<\/span><\/td>\n | 10mg<\/span><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n \n \u2014 \u2014Written\/Edited by V. Shallan\u3010\u7248\u6743\u5f52\u96c6\u5947\u751f\u7269\/MKBio\u61cb\u5eb7\u6240\u6709\u3011<\/span><\/p>\n <\/p>\n","protected":false},"excerpt":{"rendered":" \u66f4\u591a\u4ea7\u54c1\u4fe1\u606f\u8bf7\u54a8\u8be2\u5ba2\u670d\uff01<\/p>\n","protected":false},"featured_media":26239,"template":"","meta":[],"product_cat":[99],"product_tag":[105],"_links":{"self":[{"href":"\/\/www.562part.com\/wp-json\/wp\/v2\/product\/3648"}],"collection":[{"href":"\/\/www.562part.com\/wp-json\/wp\/v2\/product"}],"about":[{"href":"\/\/www.562part.com\/wp-json\/wp\/v2\/types\/product"}],"wp:featuredmedia":[{"embeddable":true,"href":"\/\/www.562part.com\/wp-json\/wp\/v2\/media\/26239"}],"wp:attachment":[{"href":"\/\/www.562part.com\/wp-json\/wp\/v2\/media?parent=3648"}],"wp:term":[{"taxonomy":"product_cat","embeddable":true,"href":"\/\/www.562part.com\/wp-json\/wp\/v2\/product_cat?post=3648"},{"taxonomy":"product_tag","embeddable":true,"href":"\/\/www.562part.com\/wp-json\/wp\/v2\/product_tag?post=3648"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}} | | | | | | | | | | | | |