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  • 2012

    10-22

    培養細(xi)(xi)(xi)(xi)胞(bao)(bao)的(de)常(chang)規檢查(cha)和觀(guan)察(cha)方法(fa)細(xi)(xi)(xi)(xi)胞(bao)(bao)培養后,需要對其(qi)生長(chang)(chang)狀況(kuang)、形(xing)態(tai)(tai)甚至生物學性狀連續(xu)地進行(xing)觀(guan)察(cha).由于(yu)細(xi)(xi)(xi)(xi)胞(bao)(bao)小而復(fu)雜,若(ruo)不借(jie)助(zhu)適當(dang)的(de)手(shou)段,則難以觀(guan)察(cha)期形(xing)態(tai)(tai)、結構,更難發現細(xi)(xi)(xi)(xi)胞(bao)(bao)內各種組(zu)分(fen)的(de)分(fen)子組(zu)成及(ji)(ji)功(gong)能.目(mu)前,已有(you)多種研究細(xi)(xi)(xi)(xi)胞(bao)(bao)的(de)技(ji)術,從光學顯微(wei)鏡到電子顯微(wei)鏡,從一般細(xi)(xi)(xi)(xi)胞(bao)(bao)化學法(fa)到免疫化學法(fa).細(xi)(xi)(xi)(xi)胞(bao)(bao)培養后,需要對其(qi)生長(chang)(chang)狀況(kuang)、形(xing)態(tai)(tai)甚至生物學性狀連續(xu)地進行(xing)觀(guan)察(cha).由于(yu)細(xi)(xi)(xi)(xi)胞(bao)(bao)小而復(fu)雜,若(ruo)不借(jie)助(zhu)適當(dang)的(de)手(shou)段,則難以觀(guan)察(cha)期形(xing)態(tai)(tai)、結構,更難發現細(xi)(xi)(xi)(xi)胞(bao)(bao)內各種組(zu)分(fen)的(de)分(fen)子組(zu)成及(ji)(ji)功(gong)能.目(mu)前,已有(you)多種研究細(xi)(xi)(xi)(xi)胞(bao)(bao)的(de)技(ji)術,從光...

  • 2012

    10-11

    上(shang)(shang)皮(pi)(pi)(pi)(pi)(pi)(pi)細(xi)(xi)胞(bao)的培養(yang)(yang)方法(fa)上(shang)(shang)皮(pi)(pi)(pi)(pi)(pi)(pi)細(xi)(xi)胞(bao)的培養(yang)(yang)方法(fa)上(shang)(shang)皮(pi)(pi)(pi)(pi)(pi)(pi)細(xi)(xi)胞(bao)包(bao)括腺上(shang)(shang)皮(pi)(pi)(pi)(pi)(pi)(pi)是(shi)(shi)很多器(qi)官如肝、胰、乳腺等的功能成分(fen),又由于(yu)癌(ai)起(qi)源于(yu)上(shang)(shang)皮(pi)(pi)(pi)(pi)(pi)(pi)組織,故上(shang)(shang)皮(pi)(pi)(pi)(pi)(pi)(pi)細(xi)(xi)胞(bao)培養(yang)(yang)特別受到重(zhong)視.但上(shang)(shang)皮(pi)(pi)(pi)(pi)(pi)(pi)細(xi)(xi)胞(bao)培養(yang)(yang)中常混雜有成纖維細(xi)(xi)胞(bao),培養(yang)(yang)時生(sheng)(sheng)(sheng)(sheng)長(chang)(chang)速度往(wang)往(wang)超過上(shang)(shang)皮(pi)(pi)(pi)(pi)(pi)(pi)細(xi)(xi)胞(bao),并難以(yi)(yi)純化,同時上(shang)(shang)皮(pi)(pi)(pi)(pi)(pi)(pi)細(xi)(xi)胞(bao)難以(yi)(yi)在體(ti)外(wai)長(chang)(chang)期(qi)生(sheng)(sheng)(sheng)(sheng)存,因此純化和延長(chang)(chang)生(sheng)(sheng)(sheng)(sheng)存時間是(shi)(shi)培養(yang)(yang)關(guan)鍵.體(ti)內上(shang)(shang)皮(pi)(pi)(pi)(pi)(pi)(pi)細(xi)(xi)胞(bao)生(sheng)(sheng)(sheng)(sheng)長(chang)(chang)在膠原(yuan)(yuan)構成的基膜,因此培養(yang)(yang)在有膠原(yuan)(yuan)的底物上(shang)(shang)可(ke)能利于(yu)生(sheng)(sheng)(sheng)(sheng)長(chang)(chang),另(ling)外(wai)人或小鼠(shu)表皮(pi)(pi)(pi)(pi)(pi)(pi)細(xi)(xi)胞(bao)培養(yang)(yang)在以(yi)(yi)3T3細(xi)(xi)胞(bao)為飼養(yang)(yang)層(用射線(xian)照射后)時,細(xi)(xi)胞(bao)易生(sheng)(sheng)(sheng)(sheng)長(chang)(chang)并可(ke)發生(sheng)(sheng)(sheng)(sheng)一(yi)定(ding)程度的分(fen)化現象.降低(di)PH、Ca...

  • 2012

    3-27

    細(xi)胞(bao)(bao)的復(fu)蘇(su)及(ji)凍存(cun)方(fang)(fang)法一.細(xi)胞(bao)(bao)復(fu)蘇(su)與培養(yang)將(jiang)液氮或-80℃保(bao)存(cun)的腫(zhong)(zhong)瘤(liu)(liu)細(xi)胞(bao)(bao)于(yu)37℃水浴,快速溶化(hua),用(yong)8.0ml培養(yang)基混(hun)勻已融化(hua)的腫(zhong)(zhong)瘤(liu)(liu)細(xi)胞(bao)(bao)懸(xuan)液,于(yu)1500轉/分,離心3分鐘(zhong)。棄上清,再吸取8.0ml培養(yang)基質混(hun)勻細(xi)胞(bao)(bao)沉淀(dian),再1500轉/分,離心3分鐘(zhong),棄上清,細(xi)胞(bao)(bao)沉淀(dian)用(yong)1.0ml培養(yang)基混(hun)勻,備(bei)用(yong)。另取一個75cm2方(fang)(fang)瓶,加入(ru)14.0ml培養(yang)基質,將(jiang)上述制備(bei)的含(han)腫(zhong)(zhong)瘤(liu)(liu)細(xi)胞(bao)(bao)懸(xuan)液(1.0ml)加入(ru)此方(fang)(fang)瓶中,于(yu)37℃,5%CO2孵(fu)育箱(xiang)中培養(yang)。若此腫(zhong)(zhong)瘤(liu)(liu)細(xi)胞(bao)(bao)懸(xuan)浮生長,大約3-4天細(xi)胞(bao)(bao)基質會(hui)變黃,5....

  • 2012

    3-15

    微生物(wu)菌種(zhong)冷凍干燥和液氮(dan)保藏菌種(zhong)技術-、安(an)瓶管(guan)開(kai)封1.用(yong)浸(jin)過70%酒精的(de)脫(tuo)脂(zhi)棉(mian)擦凈安(an)瓿管(guan)。2.用(yong)火焰將安(an)瓿管(guan)頂(ding)(ding)端加熱(re)。3.滴(di)無菌水至加熱(re)的(de)安(an)瓿管(guan)頂(ding)(ding)端使玻璃開(kai)裂(lie)。4.用(yong)挫刀或鑷子(zi)敲下已開(kai)裂(lie)的(de)安(an)瓿管(guan)的(de)頂(ding)(ding)端。二、菌株恢復培(pei)養(yang)(yang)1.用(yong)無菌吸管(guan),吸取0.3~0.5ml適(shi)(shi)宜的(de)液體(ti)(ti)培(pei)養(yang)(yang)基,滴(di)人(ren)安(an)瓿管(guan)內(nei),輕(qing)輕(qing)振(zhen)蕩,使凍于菌體(ti)(ti)溶解呈懸浮狀。2.取0.1~0.2ml菌體(ti)(ti)懸浮液,移植于適(shi)(shi)宜的(de)瓊脂(zhi)斜面/平板培(pei)養(yang)(yang)基上,剩(sheng)余的(de)菌液,注入適(shi)(shi)宜的(de)液體(ti)(ti)培(pei)養(yang)(yang)基內(nei),然后在(zai)建議的(de)溫度下培(pei)養(yang)(yang)。3.若未(wei)能(neng)活化,或活化后有...

  • 2012

    3-7

    大(da)腸桿菌一般培(pei)養方(fang)法使用牛(niu)肉膏(gao)蛋(dan)白胨(dong)培(pei)養基組成成分:牛(niu)肉膏(gao)3g,蛋(dan)白胨(dong)10g,Nacl5g,瓊脂15~20g,水1000mL,PH7.4~7.6。具體配(pei)置(zhi)步驟:1.稱(cheng)(cheng)(cheng)藥(yao)品(pin)(pin)按實際用量(liang)(liang)計算后,按配(pei)方(fang)稱(cheng)(cheng)(cheng)取各種藥(yao)品(pin)(pin)放入(ru)大(da)燒(shao)杯中.牛(niu)肉膏(gao)可放在(zai)(zai)小(xiao)燒(shao)杯或表(biao)面(mian)皿中稱(cheng)(cheng)(cheng)量(liang)(liang),用熱(re)(re)水溶(rong)解(jie)后倒入(ru)大(da)燒(shao)杯;也可放在(zai)(zai)稱(cheng)(cheng)(cheng)量(liang)(liang)紙(zhi)上稱(cheng)(cheng)(cheng)量(liang)(liang),隨后放入(ru)熱(re)(re)水中,牛(niu)肉膏(gao)使與(yu)稱(cheng)(cheng)(cheng)量(liang)(liang)紙(zhi)分離,立(li)即(ji)取出(chu)紙(zhi)片.蛋(dan)白胨(dong)極易吸潮,故稱(cheng)(cheng)(cheng)量(liang)(liang)時要迅速.2.加(jia)熱(re)(re)溶(rong)解(jie)在(zai)(zai)燒(shao)杯中加(jia)入(ru)少于(yu)所需要的水量(liang)(liang),然(ran)后放在(zai)(zai)石棉網上,小(xiao)火加(jia)熱(re)(re),并用玻棒(bang)攪拌,待(dai)藥(yao)品(pin)(pin)*溶(rong)解(jie)...

  • 2012

    2-28

    細(xi)胞(bao)(bao)傳代的一般方(fang)法開始細(xi)胞(bao)(bao)傳代前,仔(zi)細(xi)檢(jian)查細(xi)胞(bao)(bao)傳代所需(xu)的儀器(qi)和試劑(ji)是否齊(qi)全:一般主(zhu)要有:細(xi)胞(bao)(bao)(單層細(xi)胞(bao)(bao),鏡檢(jian)適合)、培養(yang)液(ye)(ye)(MEM-0.2%LH培養(yang)液(ye)(ye)或MEM培養(yang)液(ye)(ye)或199等適合細(xi)胞(bao)(bao)有要求(qiu)的培養(yang)液(ye)(ye))、滅活小牛血清、慶大(da)霉素溶液(ye)(ye)(1萬(wan)單位(wei))、7.5%NaHC03溶液(ye)(ye)、0.25%胰(yi)蛋(dan)白酶、PBS洗液(ye)(ye)。用具(ju):小方(fang)瓶(100m1)、克氏瓶、膠塞、吸管(10ml、1m1)、細(xi)胞(bao)(bao)吹打管(20ml)(以上用具(ju)需(xu)經121℃至少(shao)15分鐘(zhong)高壓(ya)滅菌(jun))C02孵(fu)箱(xiang)、超凈臺、倒置顯微鏡、4℃、—20℃冰...

  • 2012

    2-23

    一、常規操作1.穿著(zhu)實驗服。自己的(de)(de)白(bai)大衣或厚(hou)外套(tao),可脫在(zai)門(men)外的(de)(de)衣帽架上。2.穿著(zhu)拖鞋(xie)。在(zai)緩(huan)沖間(jian)(jian)換下自己的(de)(de)鞋(xie)子,盡(jin)量避(bi)免在(zai)緩(huan)沖間(jian)(jian)走動、停留。3.細胞(bao)房zui多可4個人同時(shi)使(shi)用(yong)(yong)。盡(jin)量避(bi)免在(zai)內長時(shi)間(jian)(jian)逗留、交談。二、培(pei)養(yang)箱使(shi)用(yong)(yong)規則(ze)1.從培(pei)養(yang)箱取(qu)放(fang)物(wu)品前,用(yong)(yong)酒(jiu)精清(qing)潔(jie)雙手(shou)(shou)(或手(shou)(shou)套(tao)),盡(jin)量縮(suo)短開門(men)時(shi)間(jian)(jian)和減少開門(men)次數。注:培(pei)養(yang)箱中的(de)(de)空氣(qi)是經過過濾的(de)(de)潔(jie)凈空氣(qi)。長時(shi)間(jian)(jian)敞開或頻繁開關培(pei)養(yang)箱,容易造成污染。2.培(pei)養(yang)瓶皿(min)放(fang)入培(pei)養(yang)箱前,用(yong)(yong)酒(jiu)精消毒表(biao)面,并稍等(deng)至酒(jiu)精揮發后再放(fang)入,以免培(pei)養(yang)箱內滯留過多乙醇蒸...

  • 2011

    12-15

    年末抗(kang)(kang)體(ti)(ti)*啦!年末抗(kang)(kang)體(ti)(ti)*啦!為回饋廣大客戶(hu)我公(gong)(gong)(gong)司(si)(si)目前有(you)以下抗(kang)(kang)體(ti)(ti),*,價格低廉(lian),保證質量!2鼠抗(kang)(kang)人Ki-67(單(dan)克(ke)隆(long)(long)),公(gong)(gong)(gong)司(si)(si):LDako,兔單(dan)克(ke)隆(long)(long)ErbB2/Her-2抗(kang)(kang)體(ti)(ti),公(gong)(gong)(gong)司(si)(si):Eptitomics鼠抗(kang)(kang)雌激(ji)素受(shou)體(ti)(ti)(EstrogenReceptor)單(dan)克(ke)隆(long)(long)抗(kang)(kang)體(ti)(ti),公(gong)(gong)(gong)司(si)(si):Beckmancoulter,鼠抗(kang)(kang)Glypican-3單(dan)克(ke)隆(long)(long)抗(kang)(kang)體(ti)(ti),公(gong)(gong)(gong)司(si)(si):Bio-SB抗(kang)(kang)NCL-HPP-A幽門桿菌抗(kang)(kang)體(ti)(ti)(多克(ke)隆(long)(long)),公(gong)(gong)(gong)司(si)(si):Novocastra,下面是來自哈佛(fo)實驗室的抗(kang)(kang)體(ti)(ti):兔抗(kang)(kang)鼠肥大細胞抗(kang)(kang)體(ti)(ti):mmcp-...

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