【摘要】：第一部分 納米細(xì)菌的培養(yǎng)、收獲與鑒定 目的：探討納米細(xì)菌的培養(yǎng)和收獲方法，檢測(cè)培養(yǎng)過(guò)程中物質(zhì)消耗情況和生長(zhǎng)速度變化，鑒定培養(yǎng)后的納米細(xì)菌。 方法：在納米細(xì)菌的培養(yǎng)過(guò)程中，尋找培養(yǎng)的適宜條件，防污染及收獲方法。用全自動(dòng)生化儀動(dòng)態(tài)檢測(cè)培養(yǎng)上清液中電解質(zhì)和葡萄糖的變化情況。使用酶標(biāo)儀動(dòng)態(tài)檢測(cè)納米細(xì)菌的吸光度值，間接了解其生長(zhǎng)速度。通過(guò)使用納米細(xì)菌單克隆抗體進(jìn)行免疫熒光試驗(yàn)鑒定培養(yǎng)的納米細(xì)菌和利用電鏡進(jìn)一步觀察其形狀。 結(jié)果：納米細(xì)菌適宜的培養(yǎng)條件是：使用細(xì)胞培養(yǎng)方法，含10％FBS的完全培養(yǎng)基，加入納米細(xì)菌后，在培養(yǎng)箱中培養(yǎng)，培養(yǎng)箱設(shè)置為37℃，5％CO2和95％空氣培養(yǎng)。培養(yǎng)過(guò)程中采用一系列措施可有效地避免納米細(xì)菌污染。培養(yǎng)的納米細(xì)菌用細(xì)胞刮子刮離瓶底，吸入離心管中，4℃離心45分鐘，將細(xì)菌粉末-20℃保存，可以較好地收獲納米細(xì)菌。培養(yǎng)過(guò)程中，上清液中的K、Na、Cl、Mg及葡萄糖的濃度無(wú)明顯變化(P＞0.05)，而Ca、P進(jìn)行性減少，每周檢測(cè)結(jié)果比較差異有顯著性(P＜0.05)。納米細(xì)菌吸光度動(dòng)態(tài)檢測(cè)結(jié)果顯示，其吸光度值逐漸增加，每周檢測(cè)結(jié)果比較差異有顯著性(P＜0.05)。使用免疫熒光的方法檢測(cè)時(shí)，，納米細(xì)菌可發(fā)出綠色熒光，透視電鏡下測(cè)量納米細(xì)菌大小為80—300nm。
[Abstract]:The first part of the culture, harvest and identification of nanobacteria purpose: to explore the cultivation and harvesting methods of nanobacteria, The changes of substance consumption and growth rate during culture were detected to identify the cultured nanobacteria. Methods: in the process of nanobacteria culture, the suitable conditions, pollution prevention and harvest methods were found. The changes of electrolyte and glucose in culture supernatant were detected by automatic biochemical instrument. The absorbance value of nanobacteria was dynamically detected by enzyme-labeled instrument, and the growth rate of nanobacteria was indirectly understood. The cultured nanobacteria were identified by immunofluorescence assay using monoclonal antibodies against nanobacteria and their shapes were observed by electron microscope. Results: the suitable culture conditions for nanobacteria were as follows: using cell culture method, complete culture medium containing 10%FBS, adding nanobacteria, cultured in incubator, the incubator was set at 37 鈩

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