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  • 2020

    6-15

    菌(jun)(jun)(jun)種是從事微生(sheng)物學及生(sheng)命科學研究的(de)(de)基本(ben)材料,在醫(yi)學領域中,診斷制品的(de)(de)制備,菌(jun)(jun)(jun)苗的(de)(de)生(sheng)產、微生(sheng)物致病性研究。1、梭氏法將容有1滴細胞懸液的(de)(de)小(xiao)管(guan)﹐置于盛有氫氧化(hua)鉀或(huo)五氧化(hua)二磷吸水劑的(de)(de)大試(shi)管(guan)中﹐用真(zhen)空(kong)(kong)泵抽至1、3帕時﹐將大試(shi)管(guan)密封保(bao)(bao)存。這(zhe)是為(wei)減少(shao)細胞死亡率(lv)而采(cai)取的(de)(de)一(yi)種不經(jing)凍結(jie)﹑緩(huan)慢脫水的(de)(de)干(gan)燥保(bao)(bao)藏法﹐適用于多種細菌(jun)(jun)(jun)﹑真(zhen)菌(jun)(jun)(jun)。2、冷凍真(zhen)空(kong)(kong)干(gan)燥法將細胞懸液每0、1~0、2毫(hao)升注入一(yi)無菌(jun)(jun)(jun)安(an)瓿瓶﹐于-40℃預(yu)凍1小(xiao)時﹐再于-20~30℃﹑真(zhen)空(kong)(kong)度為(wei)13、3帕的(de)(de)條件下脫水。在脫水過(guo)程(cheng)后(hou)期﹐安(an)瓿瓶外溫...

  • 2020

    4-20

    所有體(ti)外培(pei)(pei)(pei)養(yang)(yang)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao),包(bao)括初代(dai)培(pei)(pei)(pei)養(yang)(yang)及(ji)各(ge)種(zhong)(zhong)(zhong)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)系,當(dang)生長達到(dao)一定(ding)密度(du)(du)后,都需做傳(chuan)代(dai)處(chu)理(li)。傳(chuan)代(dai)的(de)頻率或間(jian)隔與培(pei)(pei)(pei)養(yang)(yang)液(ye)的(de)性質、接種(zhong)(zhong)(zhong)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)數(shu)(shu)量(liang)和(he)(he)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)增(zeng)殖速(su)度(du)(du)等有關(guan)。接種(zhong)(zhong)(zhong)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)數(shu)(shu)量(liang)大(da)、細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)基數(shu)(shu)大(da)、相同(tong)增(zeng)殖速(su)度(du)(du)條(tiao)件下,細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)數(shu)(shu)量(liang)增(zeng)加與飽和(he)(he)速(su)度(du)(du)相對要快(實際上(shang)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)接種(zhong)(zhong)(zhong)數(shu)(shu)量(liang)大(da)時(shi)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)增(zeng)殖速(su)度(du)(du)比稀少(shao)時(shi)要快)。連續細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)系和(he)(he)腫瘤細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)系比初代(dai)培(pei)(pei)(pei)養(yang)(yang)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)增(zeng)殖快,培(pei)(pei)(pei)養(yang)(yang)液(ye)中(zhong)血(xue)清含量(liang)多時(shi)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)增(zeng)殖比少(shao)時(shi)快。以上(shang)情況都會縮短傳(chuan)代(dai)時(shi)間(jian)。所謂細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)“一代(dai)”一詞,系僅指從(cong)細(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)接種(zhong)(zhong)(zhong)到(dao)分(fen)離再培(pei)(pei)(pei)養(yang)(yang)時(shi)的(de)一段時(shi)間(jian),這已成為培(pei)(pei)(pei)養(yang)(yang)工作(zuo)中(zhong)的(de)一種(zhong)(zhong)(zhong)習慣說...

  • 2020

    4-11

    細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)株(zhu)(zhu)和細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)系(xi)的(de)區別,列表說明我來答(da)這么龍厄爾尼諾LV.12019-02-24聊聊細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)株(zhu)(zhu)和細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)系(xi)的(de)區別:一、概念不(bu)同(tong)(tong)1、細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)株(zhu)(zhu):細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)株(zhu)(zhu)是通(tong)過選擇(ze)法或(huo)克(ke)隆形(xing)(xing)成法從原代培養物或(huo)細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)系(xi)中(zhong)獲得(de)具(ju)有特殊(shu)性(xing)質(zhi)或(huo)標(biao)志物的(de)培養物稱為細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)株(zhu)(zhu)(CellStrain)。2、細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)系(xi):細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)系(xi)(cellline)指原代細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)培養物經(jing)*傳代成功后所繁殖(zhi)的(de)細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)群(qun)體。也指可長期(qi)連續傳代的(de)培養細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)。二、形(xing)(xing)成方法不(bu)同(tong)(tong)1、細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)株(zhu)(zhu):通(tong)過選擇(ze)法或(huo)克(ke)隆形(xing)(xing)成法從原代培養物或(huo)細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)系(xi)中(zhong)獲得(de)具(ju)有特殊(shu)性(xing)質(zhi)或(huo)標(biao)志物的(de)培養物成為細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(bao)株(zhu)(zhu)。...

  • 2020

    4-7

    細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)在體(ti)外(wai)進行(xing)培養(yang)時(shi)影(ying)響生(sheng)長的(de)(de)(de)幾點因(yin)素細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)在體(ti)外(wai)進行(xing)培養(yang),失去了機體(ti)的(de)(de)(de)調節和控(kong)制,因(yin)此,除滿足營養(yang)的(de)(de)(de)要求外(wai),還必須使細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)生(sheng)存(cun)環(huan)(huan)境(jing)(jing)晝接近活(huo)體(ti)的(de)(de)(de)環(huan)(huan)境(jing)(jing).外(wai)環(huan)(huan)境(jing)(jing)的(de)(de)(de)培養(yang)條件如溫(wen)度(du)(du)、滲透壓、酸堿度(du)(du)等均能(neng)影(ying)響細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)的(de)(de)(de)生(sheng)長.一、溫(wen)度(du)(du):一般哺乳類及(ji)禽類細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)體(ti)外(wai)培養(yang)的(de)(de)(de)適宜溫(wen)度(du)(du)是37~38℃.溫(wen)度(du)(du)過(guo)高或過(guo)低(di)(di)都(dou)會影(ying)響到細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)的(de)(de)(de)生(sheng)長.細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)耐受低(di)(di)溫(wen)的(de)(de)(de)能(neng)力(li)比抗熱的(de)(de)(de)能(neng)力(li)強(qiang),在低(di)(di)溫(wen)下,細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)的(de)(de)(de)代謝活(huo)力(li)及(ji)核分裂降低(di)(di).溫(wen)度(du)(du)低(di)(di)于0℃時(shi),雖(sui)影(ying)響細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)代謝,但并無傷害作用.把細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)置于23~25℃時(shi),細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)仍(reng)能(neng)生(sheng)存(cun)和生(sheng)長,但速度(du)(du)減(jian)...

  • 2020

    3-19

    什么時候需要穩(wen)定細胞(bao)株?以(yi)下實驗,構建(jian)穩(wen)定細胞(bao)株而(er)不是瞬(shun)時轉(zhuan)染(ran)(ran)更能滿足實驗要求:1)外(wai)源(yuan)片段(duan)在(zai)分(fen)(fen)裂細胞(bao)中長期(qi)(2周)穩(wen)定表(biao)(biao)(biao)達(da)(da)。雖然普通轉(zhuan)染(ran)(ran)實驗一般只能保證(zheng)2-4天的轉(zhuan)染(ran)(ran)和表(biao)(biao)(biao)達(da)(da)效(xiao)(xiao)率,但腺(xian)病(bing)毒載體在(zai)多數分(fen)(fen)裂細胞(bao)中可(ke)以(yi)維持2-4周內的高(gao)轉(zhuan)染(ran)(ran)效(xiao)(xiao)率和表(biao)(biao)(biao)達(da)(da)周期(qi)。而(er)非分(fen)(fen)裂細胞(bao),可(ke)以(yi)使用腺(xian)相關(guan)病(bing)毒載體轉(zhuan)導外(wai)源(yuan)基因(yin),因(yin)為腺(xian)相關(guan)病(bing)毒載體在(zai)許多非分(fen)(fen)裂細胞(bao)中可(ke)以(yi)保持長達(da)(da)1年的高(gao)效(xiao)(xiao)表(biao)(biao)(biao)達(da)(da)。2)需要構建(jian)使用誘導表(biao)(biao)(biao)達(da)(da)系統,這主要是由于(yu):a)過表(biao)(biao)(biao)達(da)(da)基因(yin)或者干擾(rao)目(mu)的基因(yin)引起(qi)細胞(bao)毒性或者抑制(zhi)細胞(bao)生長等不利因(yin)素(su);b...

  • 2020

    3-18

    細(xi)(xi)胞(bao)培養注意事項(入門級)總的原則:無(wu)菌操作、保證細(xi)(xi)胞(bao)培養環境的穩(wen)定(ding)性按(an)時間順序整(zheng)理1、細(xi)(xi)胞(bao)房和(he)生物(wu)安全(quan)柜(ju)(或(huo)超凈(jing)工作臺(tai)(tai))先開(kai)紫(zi)外(wai)照(zhao)射30min。作用(yong):對環境滅菌(空氣)。(備注:如果著急,至(zhi)少也要開(kai)15分鐘)在做(zuo)實驗之(zhi)前考慮好需(xu)要哪些物(wu)品。開(kai)始(shi)照(zhao)射之(zhi)前,將所需(xu)要用(yong)到的物(wu)品都放(fang)(fang)到生物(wu)安全(quan)柜(ju)(或(huo)超凈(jing)工作臺(tai)(tai))里面。常(chang)用(yong)移液槍或(huo)電(dian)動移液器等,需(xu)要在工作臺(tai)(tai)上(shang)放(fang)(fang)置(zhi)一套(tao)設備。細(xi)(xi)胞(bao)培養箱一般(ban)都已經調(diao)整(zheng)好。定(ding)期留(liu)意一下二氧(yang)化碳是否足夠。不夠及時更(geng)換。2、顯微鏡(jing)需(xu)要定(ding)期消毒酒精擦拭(shi)。注意:顯微...

  • 2020

    3-9

    細(xi)(xi)(xi)胞(bao)(bao)(bao)養(yang)不好(hao)?那可(ke)能是(shi)這些沒注(zhu)意到位(wei)1、購(gou)買的(de)(de)細(xi)(xi)(xi)胞(bao)(bao)(bao)死(si)亡或(huo)存活率不佳(jia)可(ke)能原因(yin)?細(xi)(xi)(xi)胞(bao)(bao)(bao)冷(leng)凍(dong)管解凍(dong)后(hou),為(wei)何(he)會有(you)細(xi)(xi)(xi)胞(bao)(bao)(bao)數目(mu)太(tai)少的(de)(de)情形(xing)?研究(jiu)人員在細(xi)(xi)(xi)胞(bao)(bao)(bao)培養(yang)時出(chu)(chu)現存活率不佳(jia),常見原因(yin)可(ke)歸納為(wei):培養(yang)基使(shi)(shi)用(yong)錯誤(wu)或(huo)培養(yang)基品質不佳(jia);血清(qing)使(shi)(shi)用(yong)錯誤(wu)或(huo)血清(qing)的(de)(de)品質不佳(jia);解凍(dong)過程錯誤(wu);冷(leng)凍(dong)細(xi)(xi)(xi)胞(bao)(bao)(bao)解凍(dong)后(hou),加以(yi)洗滌(di)細(xi)(xi)(xi)胞(bao)(bao)(bao)和離(li)(li)心(xin);懸浮細(xi)(xi)(xi)胞(bao)(bao)(bao)誤(wu)認為(wei)死(si)細(xi)(xi)(xi)胞(bao)(bao)(bao);培養(yang)溫(wen)度使(shi)(shi)用(yong)錯誤(wu);細(xi)(xi)(xi)胞(bao)(bao)(bao)置于–80℃太(tai)久。研究(jiu)人員在冷(leng)凍(dong)細(xi)(xi)(xi)胞(bao)(bao)(bao)培養(yang)時出(chu)(chu)現細(xi)(xi)(xi)胞(bao)(bao)(bao)數目(mu)太(tai)少,大都是(shi)因(yin)為(wei)離(li)(li)心(xin)過程操作上的(de)(de)失誤(wu),造成(cheng)細(xi)(xi)(xi)胞(bao)(bao)(bao)的(de)(de)物理性損傷,以(yi)及細(xi)(xi)(xi)胞(bao)(bao)(bao)流(liu)失。建議細(xi)(xi)(xi)胞(bao)(bao)(bao)解凍(dong)后(hou)不要立刻...

  • 2020

    2-25

    細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)株(zhu)(zhu)在生(sheng)物(wu)(wu)制藥中使用非常廣泛,細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)株(zhu)(zhu)培(pei)(pei)養條件簡單,貼壁(bi)強度(du)(du)適(shi)中,比較容易轉(zhuan)染(ran),很適(shi)合用它來研究一般(ban)哺乳動物(wu)(wu)基因的功(gong)能。細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)培(pei)(pei)養過程(cheng)中的污染(ran)不僅僅指微生(sheng)物(wu)(wu),而且還包(bao)括所有混入培(pei)(pei)養環境中的、對細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)生(sheng)存(cun)有害或造成細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)不純的物(wu)(wu)質(zhi),包(bao)括生(sheng)物(wu)(wu)和化學物(wu)(wu)質(zhi)。細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)株(zhu)(zhu)的篩(shai)選我們要(yao)注意加藥時間以及維持(chi)濃度(du)(du):1.由(you)于基因轉(zhuan)染(ran)到細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)內(nei)之后要(yao)一段(duan)時間才能表達出蛋白質(zhi)。所以細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)株(zhu)(zhu)篩(shai)選時不能太(tai)早;但是也不能太(tai)晚,因為轉(zhuan)染(ran)了(le)外源基因的細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)代(dai)謝負荷(he)較大,增值較慢,時間長了(le)就(jiu)會被沒有外源基因轉(zhuan)入的細(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)所淹(yan)沒,終...

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