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”的de最zui可ke靠kao的de方fang法fa之zhi一yi。但dan操cao作zuo技ji術shu要yao求qiu高gao,在zai顯xian微wei鏡jing下xia,先xian用yong豬zhu眼yan瞼jian毛mao做zuo成cheng的de解jie剖pou針zhen,將jiang水shui滴di中zhong雜za物wu和he不bu需xu要yao的de藻zao細xi胞bao,撥bo向xiang一yi邊bian,將jiang所suo需xu的de藻zao細xi胞bao撥bo向xiang另ling一yi邊bian;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9xiaokuai,xijinglianggan,zhuangzaipeiyangminzhong,ganzaomiejun,bazhexiexiaokuaibopianpailiezaimiejundezaibopianshang,zaimeikuaixiaobopianshang,diyixiaodihanyouweizaodeshuiyang,shuididedaxiao,yizaixianweijingdibeijingxianengkanqingshuididequanbuhuodabufen,shaoshaoyidongxianweijingdezaiwutai,jiunengkanqingzhenggexiaoshuidiweidu,faxianxiaoshuidizhongzhiyouyigesuoyaofenlidezaoxibao,wuqitashengwuhunzashi,jiukeyongxiaoduniezi,jianggaibopianjiaqilai,fangjinzhuangyoupeiyangyedexiaoshiguanzhongjinxingpeiyang。ruguobushiweilededao“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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