- MTS细胞增殖检测方法
- Addexbio 品牌介绍及代理商/经销商名单品牌专区 .....
- NovateinBio代理商名录品牌:NovateinBio...
- 大鼠神经肽Y(NPY)ELISA试剂盒使用说明书_仪表网
- SMCGlobal Network South Korea
- 下列不属于消化酶抑制剂的是a胰蛋白酶抑制剂b胰凝乳蛋白酶抑制...
- 即用型肿瘤细胞冻存液操作步骤
- Vectorlabs H1200原装H1200 VECTAS...
- 湖南都市职业学院2020、2019各省录取分数线|专业录取分...
- 我国兽药产业的现状与发展趋势_企业
- 关于恒温核酸扩增检测仪等22个产品分类界定的通知杭州科人医药...
- AGS细胞专用培养基
- [07-20]进口细胞株,进口细胞株厂家.进口细胞株价格上海谷研实业有限公司
- [10-06]精液中有圆细胞是什么意思呢真实医生回答春雨医生
- [07-24]【求助】不同肝癌细胞系之间的比较 细胞技术讨论版论坛
- [10-02]尿检白细胞1+是什么意思?_39健康问答_
- [07-22]人骨髓瘤细胞系KGHM的建立
- [10-02]尿常规检查中鳞状上皮细胞偏高是怎么回事__
- [10-02]尿常规中的上皮细胞正常值多少个?_有问必答_
- [10-02]尿检白细胞和上皮细胞高
- [10-01]【尿常规里出现小圆细胞阳性是怎么回事?】_怎样_如何 大众养生网
Freezing and Thawing cells
蚂蚁淘
/发布时间:1545761421 /浏览量:86
FreezingandThawingcells
Freezing
Itisbesttofreezecellsthataregrowingrapidly.Withadherentcells,itiseasiesttosetup100mmdishes,givethemfreshmediumthedaybeforeyoufreezethem,andfreezethemjustastheybecomeconfluent.IfyouareworkingwithsUSPensioncells,makesurethattheyaregrowingvigorouslybeforeyoufreezethem.1.Trypsinizethecellsifnecessary,andthenspinthemdown.Resuspendtheminonehalfthevolumethatyouwanttoendupwith.
Inotherwords,use0.5mlper100mmdishyouareusing.
ThecellsshouldberesuspendedinDMEMcontaining20%fetalcalfserum.Putthetubeinanicebucketandletthecellscool.
2.Now,addtheDMSOtothecellsslowly.ThisstepisaccomplishedbyaddingtotheresuspendedcellsanequalvolumeofDMEMcontaining20%fetalcalfserumand20%DMSO.
ThereasontodothisslowlyistoavoidosmoticshockandtominimizetheheatingthatoccurswhenDMSOisaddedtowater.Whenyouarefinished,youshouldhavethecellssuspendedinDMEMcontaining20%fetalcalfserumand10%DMSOwithallthecellsfromasingle100mmdishin1ml.
3.Chillthecellsonice,inthecentrifugetube,forexactly30min.Thentransferthemtofreezingvialswhichhavebeenpre-chilledinthefreezerintheplasticrack,whichhasalsobeenpre-chilledinthefreezer.Use1mlpervial.4.Putthevialsinawellinsulatedboxthathasalsobeenpre-chilledinthefreezer.Insulationisbestaccomplishedwithcotton.Thenputtheboxinthe-70°Cfreezerovernight,forslowcooling,andthenputthemintoliquidnitrogen.Cellswillstoreforeverinliquidnitrogen.Storageat-70°Cisriskier.
Cellsshouldbethawedrapidlyandthendilutedslowlyintowarmgrowthmedium.
Thenputthecellsona100mldish.Thiswaythecellsdon"tsufferosmoticshock.
Somecellsseemtobenefitfrombeingdilutedintomedium,spundown,andthenresuspendedinwarmmedium.ThisgetsridoftheDMSO,butmaydisturbfragilecells.Ifthecellsareseededwithoutspinning,themediumshouldbechangedassoonasthecellshaveattached.
================ 蚂蚁淘在线 ================
免责声明：本文仅代表作者个人观点，与本网无关。其创作性以及文中陈述文字和内容未经本站证实，对本文以及其中全部或者部分内容、文字的真实性、完整性、及时性本站不做任何保证或承诺，请读者仅作参考，并请自行核实相关内容
版权声明：未经蚂蚁淘在线授权不得转载、摘编或利用其他方式使用上述作品。已经经本网授权使用作品的，应该授权范围内使用，并注明“来源：蚂蚁淘在线”。违反上述声明者，本网将追究其相关法律责任。
