培養微藻,首先要從天然海水中分離出所需要的“單種”或“克隆”,如果在培養過程中,一旦發生雜藻汙染或遭到蟲害。必須重新進行藻種的分離,以保證藻種具備“單種”或“克降”的特性。
微藻培養者根據各自的經驗和習慣,創造了各式各樣的藻種分離篩選方法,已有十幾種之多。最常用的方法是微細吸管法、小水滴法和瓊脂平板法。
微細吸管法最常用,是取得“單種”或“克隆”dezuikekaodefangfazhiyi。dancaozuojishuyaoqiugao,zaixianweijingxia,xianyongzhuyanjianmaozuochengdejiepouzhen,jiangshuidizhongzawuhebuxuyaodezaoxibao,boxiangyibian,jiangsuoxudezaoxibaoboxianglingyibian;對dui於yu有you趨qu光guang性xing的de鞭bian毛mao藻zao,可ke在zai一yi側ce投tou入ru光guang線xian,則ze所suo要yao選xuan的de鞭bian毛mao藻zao遊you向xiang有you光guang一yi側ce,雜za質zhi留liu在zai原yuan處chu,這zhe樣yang用yong玻bo璃li微wei細xi吸xi管guan吸xi取qu所suo需xu的de藻zao細xi胞bao就jiu比bi較jiao容rong易yi,另ling外wai,對dui個ge體ti較jiao大da的de種zhong類lei,可ke在zai雙shuang筒tong解jie剖pou鏡jing下xia操cao作zuo,將jiang微wei細xi吸xi管guan內nei的de藻zao細xi胞bao輕qing輕qing吹chui入ru淺qian凹ao栽zai玻bo片pian上shang或huo栽zai玻bo片pian上shang的de水shui滴di中zhong,為wei達da到dao純chun培pei養yang,微wei藻zao細xi胞bao最zui好hao經jing數shu次ci洗xi滌di,即ji吹chui入ru另ling一yi滴di之zhi後hou,再zai吸xi出chu吹chui入ru另ling一yi滴di滅mie菌jun培pei養yang液ye水shui滴di中zhong,反fan複fu稱cheng出chu洗xi滌di幾ji次ci,最zui後hou經jing鏡jing檢jian確que認ren水shui滴di中zhong是shi要yao篩shai選xuan的de一yi個ge藻zao細xi胞bao之zhi後hou,再zai接jie種zhong試shi管guan中zhong,或huo將jiang含han有you藻zao細xi胞bao的de一yi段duan毛mao細xi管guan用yong消xiao毒du鑷nie子zi夾jia斷duan,放fang入ru接jie種zhong試shi管guan中zhong。
小水滴法比較方便易行,先將24乘24毫米的蓋玻片,用割針劑安瓿的小砂輪分割成8乘8毫米的9小xiao塊kuai,洗xi淨jing晾liang幹gan,裝zhuang在zai培pei養yang皿min中zhong,幹gan燥zao滅mie菌jun,把ba這zhe些xie小xiao塊kuai玻bo片pian排pai列lie在zai滅mie菌jun的de栽zai玻bo片pian上shang,在zai每mei塊kuai小xiao玻bo片pian上shang,滴di一yi小xiao滴di含han有you微wei藻zao的de水shui樣yang,水shui滴di的de大da小xiao,以yi在zai顯xian微wei鏡jing低di倍bei鏡jing下xia能neng看kan清qing水shui滴di的de全quan部bu或huo大da部bu分fen,稍shao稍shao移yi動dong顯xian微wei鏡jing的de栽zai物wu台tai,就jiu能neng看kan清qing整zheng個ge小xiao水shui滴di為wei度du,發fa現xian小xiao水shui滴di中zhong隻zhi有you一yi個ge所suo要yao分fen離li的de藻zao細xi胞bao,無wu其qi他ta生sheng物wu混hun雜za時shi,就jiu可ke用yong消xiao毒du鑷nie子zi,將jiang該gai玻bo片pian夾jia起qi來lai,放fang進jin裝zhuang有you培pei養yang液ye的de小xiao試shi管guan中zhong進jin行xing培pei養yang。如ru果guo不bu是shi為wei了le得de到dao“克隆”,水滴中隻要是同一種藻細胞,2-3個細胞成鏈也行,這樣更易使所需要的藻類迅速生長繁殖起來。
瓊脂平板法對分離能運動的羽紋矽藻類、鞭毛型綠藻類最為適宜,方法易於掌握。這又可分為劃線接種法、噴霧接種法和小水滴擴散法。現僅舉小水滴擴散法為例。在製成的1-1.5%瓊脂,加熱攪拌,稍冷後分裝於培養皿中,厚3-5毫米,經高壓滅菌,放冷)上(shang),稀(xi)疏(shu)地(di)滴(di)上(shang)一(yi)些(xie)水(shui)樣(yang)小(xiao)滴(di),放(fang)入(ru)培(pei)養(yang)箱(xiang)中(zhong)或(huo)北(bei)窗(chuang)口(kou)下(xia)培(pei)養(yang),當(dang)小(xiao)淼(miao)貯(zhu)存(cun)器(qi)的(de)藻(zao)類(lei)隨(sui)水(shui)滴(di)的(de)擴(kuo)散(san)和(he)藻(zao)類(lei)自(zi)身(shen)的(de)運(yun)動(dong),分(fen)散(san)成(cheng)單(dan)個(ge)個(ge)體(ti),在(zai)某(mou)些(xie)方(fang)麵(mian)平(ping)板(ban)的(de)某(mou)一(yi)部(bu)位(wei)形(xing)成(cheng)藻(zao)落(luo),在(zai)顯(xian)微(wei)鏡(jing)下(xia)檢(jian)查(zha)確(que)認(ren)後(hou),用(yong)消(xiao)毒(du)過(guo)的(de)玻(bo)璃(li)微(wei)針(zhen)取(qu)下(xia)所(suo)需(xu)藻(zao)落(luo),放(fang)入(ru)試(shi)管(guan)中(zhong)培(pei)養(yang),反(fan)複(fu)幾(ji)次(ci),就(jiu)能(neng)得(de)到(dao)純(chun)種(zhong)培(pei)養(yang)物(wu)。
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