含有一種以上的微生物培養物稱為混和培養物(mixed culture)。zaizhenduimouxiexijundejiancexiangmuli,yinweizaibiaoyangzhongchangchanghunyouduozhongjun,womentongchangxuyaozhenduimouyimudejunjinxingfenlichunhua。tongguofenlichunhua,womennenggoulianghaodipinggugaibiaoyangdeweishengwuzhibiaoshuju,changchangyongyuwuranqingkuangjianding、活性物質檢測、產品質量參數評估等。如果在一個菌落中所有細胞均來自於一個親代細胞,那麼這個菌落稱為純培養(pureculture)。在進行菌種鑒定時,所用的微生物一般均要求為純的培養物。得到純培養的過程稱為分離純化,主要方法有以下六種。
1.傾注平板法
首先把微生物懸液通過一係列稀釋,取一定量的稀釋液與熔化好的保持在40-50°Czuoyoudeyingyangqiongzhipeiyangjichongfenhunhe,ranhoubazhehunheyeqingzhudaowujundepeiyangminzhong,dainingguzhihou,bazhepingbandaozhizaihengxiangzhongpeiyang。danyixibaojingguoduocizengzhihouxingchengyigejunluo,qudangejunluozhichengxuanye,zhongfushangshubuzhoushuci,biankededaochunpeiyangwu。
2.塗布平板法
首shou先xian把ba微wei生sheng物wu懸xuan液ye通tong過guo適shi當dang的de稀xi釋shi,取qu一yi定ding量liang的de稀xi釋shi液ye放fang在zai無wu菌jun的de已yi經jing凝ning固gu的de營ying養yang瓊qiong脂zhi平ping板ban上shang,然ran後hou用yong無wu菌jun的de玻bo璃li刮gua刀dao把ba稀xi釋shi液ye均jun勻yun地di塗tu布bu在zai培pei養yang基ji表biao麵mian上shang,經jing恒heng溫wen培pei養yang便bian可ke以yi得de到dao單dan個ge菌jun落luo。
3.平板劃線法
zui 簡jian單dan的de分fen離li微wei生sheng物wu的de方fang法fa是shi平ping板ban劃hua線xian法fa。用yong無wu菌jun的de接jie種zhong環huan取qu培pei養yang物wu少shao許xu在zai平ping板ban上shang進jin行xing劃hua線xian。劃hua線xian的de方fang法fa很hen多duo,常chang見jian的de比bi較jiao容rong易yi出chu現xian單dan個ge菌jun落luo的de劃hua線xian方fang法fa有you斜xie線xian法fa、曲線法、方格法、放射法、四(si)格(ge)法(fa)等(deng)。當(dang)接(jie)種(zhong)環(huan)在(zai)培(pei)養(yang)基(ji)表(biao)麵(mian)上(shang)往(wang)後(hou)移(yi)動(dong)時(shi),接(jie)種(zhong)環(huan)上(shang)的(de)菌(jun)液(ye)逐(zhu)漸(jian)稀(xi)釋(shi),最(zui)後(hou)在(zai)所(suo)劃(hua)的(de)線(xian)上(shang)分(fen)散(san)著(zhe)單(dan)個(ge)細(xi)胞(bao),經(jing)培(pei)養(yang),每(mei)一(yi)個(ge)細(xi)胞(bao)長(chang)成(cheng)一(yi)個(ge)菌(jun)落(luo)。
4.富集培養法
富(fu)集(ji)培(pei)養(yang)法(fa)的(de)方(fang)法(fa)和(he)原(yuan)理(li)非(fei)常(chang)簡(jian)單(dan)。我(wo)們(men)可(ke)以(yi)創(chuang)造(zao)一(yi)些(xie)條(tiao)件(jian)隻(zhi)讓(rang)所(suo)需(xu)的(de)微(wei)生(sheng)物(wu)生(sheng)長(chang),在(zai)這(zhe)些(xie)條(tiao)件(jian)下(xia),所(suo)需(xu)要(yao)的(de)微(wei)生(sheng)物(wu)能(neng)有(you)效(xiao)地(di)與(yu)其(qi)他(ta)微(wei)生(sheng)物(wu)進(jin)行(xing)競(jing)爭(zheng),在(zai)生(sheng)長(chang)能(neng)力(li)方(fang)麵(mian)超(chao)過(guo)其(qi)他(ta)微(wei)生(sheng)物(wu)。如(ru)果(guo)要(yao)分(fen)離(li)一(yi)些(xie)專(zhuan)性(xing)寄(ji)生(sheng)菌(jun),就(jiu)必(bi)須(xu)把(ba)樣(yang)品(pin)接(jie)種(zhong)到(dao)相(xiang)應(ying)敏(min)感(gan)宿(xiu)主(zhu)細(xi)胞(bao)群(qun)體(ti)中(zhong),使(shi)其(qi)大(da)量(liang)生(sheng)長(chang)。通(tong)過(guo)多(duo)次(ci)重(zhong)複(fu)移(yi)種(zhong)便(bian)可(ke)以(yi)得(de)到(dao)純(chun)的(de)寄(ji)生(sheng)菌(jun)。
5.厭氧法
zaishiyanshizhong,weilefenlimouxieyanyangjun,keyiliyongzhuangyouyuanpeiyangjideshiguanzuoweipeiyangrongqi,bazhezhishiguanfangzaifeishuiyuzhongjiareshufenzhong,yibianzhuchupeiyangjizhongderongjieyang。ranhoukuaisulengque,bingjinxingjiezhong。jiezhonghou,jiaruwujundeshilayupeiyangjibiaomian,shipeiyangjiyukongqigejue。lingyizhongfangfashi,zaijiezhonghou,liyongN2或CO2取(qu)代(dai)培(pei)養(yang)基(ji)中(zhong)的(de)氣(qi)體(ti),然(ran)後(hou)在(zai)火(huo)焰(yan)上(shang)把(ba)試(shi)管(guan)口(kou)密(mi)封(feng)。有(you)時(shi)為(wei)了(le)更(geng)有(you)效(xiao)地(di)分(fen)離(li)某(mou)些(xie)厭(yan)氧(yang)菌(jun),可(ke)以(yi)把(ba)所(suo)分(fen)離(li)的(de)樣(yang)品(pin)接(jie)種(zhong)於(yu)培(pei)養(yang)基(ji)上(shang),然(ran)後(hou)再(zai)把(ba)培(pei)養(yang)皿(min)放(fang)在(zai)密(mi)封(feng)的(de)厭(yan)氧(yang)培(pei)養(yang)裝(zhuang)置(zhi)中(zhong)。
6.單細胞(或單孢子)分離法
是shi采cai取qu顯xian微wei分fen離li法fa從cong混hun雜za群qun體ti中zhong直zhi接jie分fen離li單dan個ge細xi胞bao或huo單dan個ge個ge體ti進jin行xing培pei養yang以yi獲huo得de純chun培pei養yang。較jiao大da的de微wei生sheng物wu,可ke采cai用yong毛mao細xi管guan提ti取qu單dan個ge個ge體ti。個ge體ti相xiang對dui較jiao小xiao的de微wei生sheng物wu,需xu采cai用yong顯xian微wei操cao作zuo儀yi,在zai顯xian微wei鏡jing下xia用yong毛mao細xi管guan或huo顯xian微wei針zhen、鉤、環等挑取單個微生物細胞或孢子以獲得純培養。單細胞分離法對操作技術有比較高的要求。
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