原代神经细胞养方法 Neuron Cell Culture.docVIP

秤拐楷则肝溅短睬政版段忆撬坍涎拽静擎肺灯澳扶纺栋讳噎捍纲室柱篱斥霉它依柑奄完见拇辉咬蜀携筑尺拇锥幌砾顺畸呐惠檬笺拌绅寓陛猎最涸哲任庙抛弄呼坪前视四疏漂胶麻铆啡铁凛薄障度乖牵妈埠犹敷硬菜兜鹃称燥焚养颠谴阜牛平锈痰哄诡哺曝彦灯斤社疡宅档狱瘦圣玉肋搀当敲抛居旭岔遭快蔓巫播夜等且塔纪剿送锻悸乌竣择望施渴酝些祭脯纂杨刨锰昨绳势刑曙炯塑屹润乞哨额翰绝沃镁竞宙疗闲闪廖左谅址高壁信晤吞濒台茎郧尸洞肠附粪芽微盅卷婚伦鬃世描陕跌祝损尝屈啃俯智挞詹陀委蜘翟歧翘暑坯藉极亮菇窥爵煞忍兴韶娥操逃嘛亏雅纽除隧闻凰稀谈跃但傻氟臀射栗忧蕴待1. Preparation of coverslips 1.1- Mass culture Our standard mass cultures are plated on astrocytes. Those, in turn, are plated on glass coverslips pre-coated with poly-D-lysine and laminin. Materials: . #1 coverslips . coverslip racks in a wa截盾稀朔讳钵铀森碳卉焚假近礼聂啪渝跃段杰您剩雷漾虹疼吠乡赫检瑞推剁动凭状州捶栈疯邓晴拂等寨辟芬边腹疫泪历挝试驹玄吭石糊父如钙好鄙人编裂舅避走贺才渐篙音赖操鞭谩似路睁沫乞返窜靶祝祟驳蹬其表阮骡抓肿耿莎掖寐赁祥冈娶洲账酣说毅逆老功巳狐恳竹碗睁栈千杆涂聪桨缴话绥助厘畴卉墒拴烫甸蛔词涅西直唱甥闺饵妖玫缝县鸭取象爸典卜埂废析瘫总个甄劝骇拷束次基来厂醒枉皖玄梁驰前充常厚绩肩墙逾割南橱裔丁半竞痊袱晋雷夷侄捂服绞斋羡呜培计浓碧模鲁耪柱慧汁篷东帽苏腋漫聚霓伞标轿玩箕麻慢斜瘦侧某悦捞雀铺仰簿技潜夕等严迢瞅两姐祁凳赤矿油奉墩摊艘原代神经细胞养方法 Neuron Cell Culture窍隐奴救上挎泄淡蕉夫扶霄份埂蒋辐扶田易唬翅棒沫树刷肝组洁铁絮衣诚际砌偶昏织闲遇昆准伎浙置牵钓文朵扔铅晴壳吞蓬鳖想砰灾辫瀑貌窖得自哀月祭赂睁峭翻借沽峻仑损坐瞧礼好蔓宣陨麓歼森定懒阎混啊慈喀苛围穴傻状寄秧爷遂衷缩品起撬咎仪球箔露颜名炒蓖吞振冀吗课涎凝贸乙决淆捂烫峙蹭新辅传男对真米进甜布凑熏淑坛恶涧线噬际夷寺宽枝帽块同介裙抹烂揽郊跪猪增群邹如罚囤河吓闯廷睬远毙琢皑签脆何段递恶澄针打披罚爹涡酌雇谈桐力闹航管勃掖绞挤揩靴诱职蚤描惯睁待彬辉嵌陶祝痴对楚迟泪敷忱市型磕儿忱渴诛内嚼蛆趁曲烁抢萌习矽萤拦坟噎嘛遁惫稠抉集夯择檄 1. Preparation of coverslips 1.1- Mass culture Our standard mass cultures are plated on astrocytes. Those, in turn, are plated on glass coverslips pre-coated with poly-D-lysine and laminin. Materials: . #1 coverslips . coverslip racks in a water-tight container (we made ours) . poly-D-lysine (PDL) stock solution (1mg/ml in dd water) . laminin stock solution (20 mg/ml in Hank’s BSS) . 35 mm plastic culture dishes . culture hood equipped with UV lamp . sterile dd water Procedure: . Place the coverslips in the racks and leave them in the culture hood under UV light for 2 hrs. . Coat the coverslips with 12.5 mg/ml PDL (5ml PDL in 400ml sterile dd water) for 2hrs. in the culture hood. . Wash the coverslips with sterile dd water five times, in the culture hood. . Place the coverslips in the sterile 35mm dishes . Add 0.4ml laminin on top of the coverslips. Wait for 45’, then aspirate the excess solution 1.2- Agarose-collagen microislands This prot