單個微生物在適宜的固體培養基表麵或內部生長、繁殖到一定程度可以形成肉眼可見的、youyidingxingtaijiegoudezixibaoshengchangqunti
，chengweijunluo。danggutipeiyangjibiaomianzhongduojunluolianchengyipianshi，bianchengweijuntai。butongweishengwuzaitedingpeiyangjishangshengchangxingchengdejunluohuojuntaiyibandoujuyouwendingdetezheng

，keyichengweiduigaiweishengwujinxingfenlei
、鑒定的重要依據。大多數細菌、酵母菌、以yi及ji許xu多duo真zhen菌jun和he單dan細xi胞bao藻zao類lei能neng在zai固gu體ti培pei養yang基ji上shang形xing成cheng孤gu立li的de菌jun落luo，采cai用yong適shi宜yi的de平ping板ban分fen離li法fa很hen容rong易yi得de到dao純chun培pei養yang。所suo謂wei平ping板ban，即ji培pei養yang平ping板ban的de簡jian稱cheng，它ta是shi指zhi固gu體ti培pei養yang基ji倒dao入ru無wu菌jun平ping皿min，冷leng卻que凝ning固gu後hou，盛sheng固gu體ti培pei養yang基ji的de平ping皿min。這zhe方fang法fa包bao括kuo將jiang單dan個ge微wei生sheng物wu分fen離li和he固gu定ding在zai固gu體ti培pei養yang基ji表biao麵mian或huo裏li麵mian。固gu體ti培pei養yang基ji用yong瓊qiong脂zhi或huo其qi它ta凝ning膠jiao物wu質zhi固gu化hua的de培pei養yang基ji，每mei個ge孤gu立li的de活huo微wei生sheng物wu體ti生sheng長chang

、繁殖形成菌落，形成的菌落便於移植。最常用的分離、培養微生物的固體培養基是瓊脂固體培養基平板。這種由Kock建立的采用平板分離微生物純培養的技術簡便易行，100多年來一直是各種菌種分離的最常用手段。

首先把微生物懸液作一係列的稀釋（如1:10、1:100、1:1000、1:10000），然後分別取不同稀釋液少許，與已熔化並冷卻至50℃左zuo右you的de瓊qiong脂zhi培pei養yang基ji混hun合he
，搖yao勻yun後hou，傾qing入ru滅mie過guo菌jun的de培pei養yang皿min中zhong，待dai瓊qiong脂zhi凝ning固gu後hou，製zhi成cheng可ke能neng含han菌jun的de瓊qiong脂zhi平ping板ban
，保bao溫wen培pei養yang一yi定ding時shi間jian即ji可ke出chu現xian菌jun落luo。如ru果guo稀xi釋shi得de當dang，在zai平ping板ban表biao麵mian或huo瓊qiong脂zhi培pei養yang基ji中zhong就jiu可ke出chu現xian分fen散san的de單dan個ge菌jun落luo，這zhe個ge菌jun落luo可ke能neng就jiu是shi由you一yi個ge細xi菌jun細xi胞bao繁fan殖zhi形xing成cheng的de。隨sui後hou挑tiao取qu該gai單dan個ge菌jun落luo，或huo重zhong複fu以yi上shang操cao作zuo數shu次ci，便bian可ke得de到dao純chun培pei養yang。

因yin為wei將jiang微wei生sheng物wu懸xuan液ye先xian加jia到dao較jiao燙tang的de培pei養yang基ji中zhong再zai倒dao平ping板ban易yi造zao成cheng某mou些xie熱re敏min感gan菌jun的de死si亡wang
，且qie采cai用yong稀xi釋shi倒dao平ping板ban法fa也ye會hui使shi一yi些xie嚴yan格ge好hao氧yang菌jun因yin被bei固gu定ding在zai瓊qiong脂zhi中zhong間jian缺que乏fa氧yang氣qi而er影ying響xiang其qi生sheng長chang，因yin此ci在zai微wei生sheng物wu學xue研yan究jiu中zhong常chang用yong的de純chun種zhong分fen離li方fang法fa是shi塗tu布bu平ping板ban法fa。其qi做zuo法fa是shi先xian將jiang已yi熔rong化hua的de培pei養yang基ji倒dao入ru無wu菌jun平ping皿min
，製zhi成cheng無wu菌jun平ping板ban，冷leng卻que凝ning固gu後hou
，將jiang一yi定ding量liang的de微wei生sheng物wu懸xuan液ye滴di加jia在zai平ping板ban表biao麵mian，再zai用yong無wu菌jun玻bo璃li塗tu棒bang將jiang菌jun液ye均jun勻yun分fen散san至zhi整zheng個ge平ping板ban表biao麵mian，經jing培pei養yang後hou挑tiao取qu單dan個ge菌jun落luo（圖1）。

最簡單的分離微生物的方法是平板劃線法，即用接種環以無菌操作沾取少許待分離的材料
，在無菌平板表麵進行連續劃線（圖2），weishengwuxibaoshuliangjiangsuizhehuaxiancishudezengjiaerjianshao，bingzhubufensankailai
，ruguohuaxianshiyidehua
，weishengwunengyiyifensan
，jingpeiyanghou
，kezaipingbanbiaomiandedaodanjunluo。youshizhezhongdanjunluobingfeidouyoudangexibaofanzhierlaide，gubixufanfufenliduocicaikededaochunzhong。qiyuanlishijiangweishengwuyangpinzaigutipeiyangjibiaomianduocizuo“由點到線”稀釋而達到分離目的的。劃線的方法很多，常見的比較容易出現單個菌落的劃線方法有斜線法

、曲線法、方格法
、放射法
、四格法等。

用yong固gu體ti培pei養yang基ji分fen離li嚴yan格ge厭yan氧yang菌jun有you特te殊shu性xing，如ru果guo該gai微wei生sheng物wu暴bao露lu於yu空kong氣qi中zhong不bu立li即ji死si亡wang，可ke以yi采cai用yong通tong常chang的de方fang法fa製zhi備bei平ping板ban
，然ran後hou置zhi放fang在zai封feng閉bi的de容rong器qi中zhong培pei養yang，容rong器qi中zhong的de氧yang氣qi可ke采cai用yong化hua學xue、物wu理li或huo生sheng物wu的de方fang法fa清qing除chu。對dui於yu那na些xie對dui氧yang氣qi更geng為wei敏min感gan的de厭yan氧yang性xing微wei生sheng物wu

，純chun培pei養yang的de分fen離li則ze可ke采cai用yong稀xi釋shi搖yao管guan培pei養yang法fa進jin行xing
，它ta是shi稀xi釋shi倒dao平ping板ban法fa的de一yi種zhong變bian通tong形xing式shi。先xian將jiang一yi係xi列lie盛sheng無wu菌jun瓊qiong脂zhi培pei養yang基ji的de試shi管guan加jia熱re使shi瓊qiong脂zhi熔rong化hua後hou冷leng卻que並bing保bao持chi在zai50℃左(zuo)右(you)，將(jiang)待(dai)分(fen)離(li)的(de)材(cai)料(liao)用(yong)這(zhe)些(xie)試(shi)管(guan)進(jin)行(xing)梯(ti)度(du)稀(xi)釋(shi)
，試(shi)管(guan)迅(xun)速(su)搖(yao)動(dong)均(jun)勻(yun)
，冷(leng)凝(ning)後(hou)
，在(zai)瓊(qiong)脂(zhi)柱(zhu)表(biao)麵(mian)傾(qing)倒(dao)一(yi)層(ceng)滅(mie)菌(jun)液(ye)體(ti)石(shi)蠟(la)和(he)固(gu)體(ti)石(shi)蠟(la)的(de)混(hun)合(he)物(wu)
，將(jiang)培(pei)養(yang)基(ji)和(he)空(kong)氣(qi)隔(ge)開(kai)。培(pei)養(yang)後(hou)，菌(jun)落(luo)形(xing)成(cheng)在(zai)瓊(qiong)脂(zhi)柱(zhu)的(de)中(zhong)間(jian)。進(jin)行(xing)單(dan)菌(jun)落(luo)的(de)挑(tiao)取(qu)和(he)移(yi)植(zhi)，需(xu)先(xian)用(yong)一(yi)隻(zhi)滅(mie)菌(jun)針(zhen)將(jiang)液(ye)體(ti)石(shi)蠟(la)--石(shi)蠟(la)蓋(gai)取(qu)出(chu)
，再(zai)用(yong)一(yi)隻(zhi)毛(mao)細(xi)管(guan)插(cha)入(ru)瓊(qiong)脂(zhi)和(he)管(guan)壁(bi)之(zhi)間(jian)
，吹(chui)入(ru)無(wu)菌(jun)無(wu)氧(yang)氣(qi)體(ti)，將(jiang)瓊(qiong)脂(zhi)柱(zhu)吸(xi)出(chu)
，置(zhi)放(fang)在(zai)培(pei)養(yang)皿(min)中(zhong)，用(yong)無(wu)菌(jun)刀(dao)將(jiang)瓊(qiong)脂(zhi)柱(zhu)切(qie)成(cheng)薄(bo)片(pian)進(jin)行(xing)觀(guan)察(cha)和(he)菌(jun)落(luo)的(de)移(yi)植(zhi)。

 

 

大(da)多(duo)數(shu)細(xi)菌(jun)和(he)真(zhen)菌(jun)
，用(yong)平(ping)板(ban)法(fa)分(fen)離(li)通(tong)常(chang)是(shi)滿(man)意(yi)的(de)，因(yin)為(wei)它(ta)們(men)的(de)大(da)多(duo)數(shu)種(zhong)類(lei)在(zai)固(gu)體(ti)培(pei)養(yang)基(ji)上(shang)長(chang)得(de)很(hen)好(hao)。然(ran)而(er)迄(qi)今(jin)為(wei)止(zhi)並(bing)不(bu)是(shi)所(suo)有(you)的(de)微(wei)生(sheng)物(wu)都(dou)能(neng)在(zai)固(gu)體(ti)培(pei)養(yang)基(ji)上(shang)生(sheng)長(chang)，例(li)如(ru)一(yi)些(xie)細(xi)胞(bao)大(da)的(de)細(xi)菌(jun)
、許多原生動物和藻類等，這些微生物仍需要用液體培養基分離來獲得純培養。

xishifashiyetipeiyangjifenlichunhuachangyongdefangfa。jiezhongwuzaiyetipeiyangjizhongjinxingshunxuxishi

，yidedaogaoduxishidexiaoguo
，shiyizhishiguanzhongfenpeibudaoyigeweishengwu。ruguojingxishihoudedaduoshushiguanzhongmeiyouweishengwushengchang，nameyouweishengwushengchangdeshiguandedaodepeiyangwukenengjiushichunpeiyangwu。ruguojingxishihoudeshiguanzhongyouweishengwushengchangdebilitigaole，dedaochunpeiyangwudejilvjiuhuijijuxiajiang。yinci
，caiyongxishifajinxingyetifenli，bixuzaitongyigexishidudexuduopingxingshiguanzhong
，daduoshu（一般應超過95%）表現為不生長。

 

 

zhinengfenlichuhunzaweishengwuquntizhongzhanshuliangyoushidezhongleishixishifadeyigezhongyaoquedian。zaiziranjie，henduoweishengwuzaihunzaquntizhongdoushishaoshu。zheshi，keyicaiquxianweifenlifaconghunzaquntizhongzhijiefenlidangexibaohuodangegetijinxingpeiyangyihuodechunpeiyang
，chengweidanxibao（或單孢子）分離法。單細胞分離法的難度與細胞或個體的大小成反比，較大的微生物如藻類、原生動物較容易
，個體很小的細菌則較難。

jiaodadeweishengwu
，kecaiyongmaoxiguantiqudangegeti，bingzaidaliangdemiejunpeiyangjizhongzhuanyiqingxijici

，chuqujiaoxiaoweishengwudewuran。zhexiangcaozuokezaidibeixianweijing，rujiepouxianweijingxiajinxing。duiyugetixiangduijiaoxiaodeweishengwu
，xucaiyongxianweicaozuoyi，zaixianweijingxiayongmaoxiguanhuoxianweizhen
、鉤、環(huan)等(deng)挑(tiao)取(qu)單(dan)個(ge)微(wei)生(sheng)物(wu)細(xi)胞(bao)或(huo)孢(bao)子(zi)以(yi)獲(huo)得(de)純(chun)培(pei)養(yang)。在(zai)沒(mei)有(you)顯(xian)微(wei)操(cao)作(zuo)儀(yi)時(shi)，也(ye)可(ke)采(cai)用(yong)一(yi)些(xie)變(bian)通(tong)的(de)方(fang)法(fa)在(zai)顯(xian)微(wei)鏡(jing)下(xia)進(jin)行(xing)單(dan)細(xi)胞(bao)分(fen)離(li)，例(li)如(ru)將(jiang)經(jing)適(shi)當(dang)稀(xi)釋(shi)後(hou)的(de)樣(yang)品(pin)製(zhi)備(bei)成(cheng)小(xiao)液(ye)滴(di)在(zai)顯(xian)微(wei)鏡(jing)下(xia)觀(guan)察(cha)，選(xuan)取(qu)隻(zhi)含(han)一(yi)個(ge)細(xi)胞(bao)的(de)液(ye)體(ti)來(lai)進(jin)行(xing)純(chun)培(pei)養(yang)物(wu)的(de)分(fen)離(li)。單(dan)細(xi)胞(bao)分(fen)離(li)法(fa)對(dui)操(cao)作(zuo)技(ji)術(shu)有(you)比(bi)較(jiao)高(gao)的(de)要(yao)求(qiu)，多(duo)限(xian)於(yu)高(gao)度(du)專(zhuan)業(ye)化(hua)的(de)科(ke)學(xue)研(yan)究(jiu)中(zhong)采(cai)用(yong)。

 

 

meiyouyizhongpeiyangjihuoyizhongpeiyangtiaojiannenggoumanzuyiqieweishengwushengchangdexuyao
，zaiyidingchengdushangsuoyoudepeiyangjidoushixuanzexingde。ruguomouzhongweishengwudeshengchangxuyaoshiyizhide，yekeyishejitedinghuanjingshizhishihezhezhongweishengwudeshengchang，yinernenggouconghunzadeweishengwuquntizhongbazhezhongweishengwuxuanzepeiyangchulai，jinguanzaihunzadeweishengwuquntizhongzhezhongweishengwukenengzhizhanshaoshu。zhezhongtongguoxuanzepeiyangjinxingweishengwuchunpeiyangfenlidejishuchengweixuanzepeiyangfenli，tebieshiyongyucongziranjiezhongfenli、xunzhaoyouyongdeweishengwu。ziranjiezhong，zaidaduoshuchangheweishengwuqunluoshiyouduozhongweishengwuzuchengde
，congzhongfenlichusuoxudetedingweishengwushishifenkunnande，youqidangmouyizhongweishengwusuocunzaideshuliangyuqitaweishengwuxiangbifeichangshaoshi，dancaiyongyibandepingbanxishifajihushibukenengde。yaofenlizhezhongweishengwu，bixugenjugaiweishengwudetedian
，baokuoyingyang
、生理、生(sheng)長(chang)條(tiao)件(jian)等(deng)，采(cai)用(yong)選(xuan)擇(ze)培(pei)養(yang)分(fen)離(li)的(de)方(fang)法(fa)。或(huo)抑(yi)製(zhi)使(shi)大(da)多(duo)數(shu)微(wei)生(sheng)物(wu)不(bu)能(neng)生(sheng)長(chang)，或(huo)造(zao)成(cheng)有(you)利(li)於(yu)該(gai)菌(jun)生(sheng)長(chang)的(de)環(huan)境(jing)，經(jing)過(guo)一(yi)定(ding)時(shi)間(jian)培(pei)養(yang)後(hou)使(shi)該(gai)菌(jun)在(zai)群(qun)落(luo)中(zhong)的(de)數(shu)量(liang)上(shang)升(sheng)，再(zai)通(tong)過(guo)平(ping)板(ban)稀(xi)釋(shi)等(deng)方(fang)法(fa)對(dui)它(ta)進(jin)行(xing)純(chun)培(pei)養(yang)分(fen)離(li)。

根據待分離微生物的特點選擇不同的培養條件，有多種方法可以采用。例如要分離高溫菌
，可在高溫條件下進行培養；要分離某種抗菌素抗性菌株，可在加有抗菌素的平板上進行分離；有些微生物如螺旋體、粘細菌、藍(lan)細(xi)菌(jun)等(deng)能(neng)在(zai)瓊(qiong)脂(zhi)平(ping)板(ban)表(biao)麵(mian)或(huo)裏(li)麵(mian)滑(hua)行(xing)，可(ke)以(yi)利(li)用(yong)它(ta)們(men)的(de)滑(hua)動(dong)特(te)點(dian)進(jin)行(xing)分(fen)離(li)純(chun)化(hua)，因(yin)為(wei)滑(hua)行(xing)能(neng)使(shi)它(ta)們(men)自(zi)己(ji)和(he)其(qi)它(ta)不(bu)能(neng)移(yi)動(dong)的(de)微(wei)生(sheng)物(wu)分(fen)開(kai)。可(ke)將(jiang)微(wei)生(sheng)物(wu)群(qun)落(luo)點(dian)種(zhong)到(dao)平(ping)板(ban)上(shang)，讓(rang)微(wei)生(sheng)物(wu)滑(hua)行(xing)，從(cong)滑(hua)行(xing)前(qian)沿(yan)挑(tiao)取(qu)接(jie)種(zhong)物(wu)接(jie)種(zhong)，反(fan)複(fu)進(jin)行(xing)，得(de)到(dao)純(chun)培(pei)養(yang)物(wu)。

富(fu)集(ji)培(pei)養(yang)法(fa)原(yuan)理(li)和(he)方(fang)法(fa)非(fei)常(chang)簡(jian)單(dan)，利(li)用(yong)不(bu)同(tong)微(wei)生(sheng)物(wu)間(jian)生(sheng)命(ming)活(huo)動(dong)特(te)點(dian)的(de)不(bu)同(tong)，製(zhi)定(ding)特(te)定(ding)的(de)環(huan)境(jing)條(tiao)件(jian)，使(shi)僅(jin)適(shi)應(ying)於(yu)該(gai)條(tiao)件(jian)的(de)微(wei)生(sheng)物(wu)旺(wang)盛(sheng)生(sheng)長(chang)，從(cong)而(er)使(shi)其(qi)在(zai)群(qun)落(luo)中(zhong)的(de)數(shu)量(liang)大(da)大(da)增(zeng)加(jia)
，很(hen)容(rong)易(yi)地(di)分(fen)離(li)到(dao)所(suo)需(xu)的(de)特(te)定(ding)微(wei)生(sheng)物(wu)。富(fu)集(ji)條(tiao)件(jian)可(ke)根(gen)據(ju)所(suo)需(xu)分(fen)離(li)的(de)微(wei)生(sheng)物(wu)的(de)特(te)點(dian)從(cong)物(wu)理(li)、化學、生物
、及綜合多個方麵進行選擇
，如溫度、pH、紫外線
、高壓、光照
、氧氣、營(ying)養(yang)等(deng)等(deng)許(xu)多(duo)方(fang)麵(mian)。在(zai)相(xiang)同(tong)的(de)培(pei)養(yang)基(ji)和(he)培(pei)養(yang)條(tiao)件(jian)下(xia)
，經(jing)過(guo)多(duo)次(ci)重(zhong)複(fu)移(yi)種(zhong)
，最(zui)後(hou)富(fu)集(ji)的(de)菌(jun)株(zhu)很(hen)容(rong)易(yi)在(zai)固(gu)體(ti)培(pei)養(yang)基(ji)上(shang)長(chang)出(chu)單(dan)菌(jun)落(luo)。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)。

 

 

fenlidemudetongchangshiyaodedaochunpeiyang。raner，zaiyouxieqingkuangxiazheshihennanzuodaode。dankeyongeryuanpeiyangwuzuoweichunhuapeiyangdetidaiwu。hanyouerzhongyishangweishengwudepeiyangwuchengweihunhepeiyangwu
，erruguopeiyangwuzhongzhihanyouerzhongweishengwu
，erqieshiyouyishidebaochierzhezhijiandetedingguanxidepeiyangwuchengweieryuanpeiyangwu。lirueryuanpeiyangwushibaocunbingdudezuiyouxiaotujing，yinweibingdushixibaoshengwudeyangedexibaoneijishengwu。youyixiejuyouxibaodeweishengwuyeshiyangedeqitashengwudexibaoneijishengwu，huoteshudegongshengguanxi。duiyuzhexieshengwu，eryuanpeiyangwushizaishiyanshikongzhitiaojianxiakenengdadaodezuijiejinyuchunpeiyangdepeiyangfangfa。lingwai，lieshixixiaoweishengwudeyuanshengdongwuyehenrongyiyongeryuanpeiyangfazaishiyanshipeiyang
，peiyangwuyouyuanshengdongwuhetalieshideweishengwuerzhezucheng。liru
，xianmaochong
、變形蟲和粘菌。