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標題: 流式質體儀:超高速之微生物鑑定定量研究 -微水珠中螢光偵測方法研究
FlowOmics: high throughput microbial identification and quantification - fluorescence detection in aqueous microdroplets
作者: 顏懷宣
Yen, Huai-Shiuan
關鍵字: 微流體生物晶片;Microfluidic biochip;微水珠形成;螢光訊號偵測;LabVIEW;Droplet;Double fluorescence;LabVIEW
出版社: 環境工程學系所
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在環境微生物與微生物生態學領域中,始終存在一個未能有效解決的問題:即在一個環境樣品中「存在微生物為何?」、「各種微生物的數量有多少?」及「細胞在做什麼?」。此方面之資訊不僅有助於解決環境微生物學與微生物生態學的基礎問題,更可應用於其他各種與微生物相關之領域,如生物合成、製藥、致病性菌株鑑定、食品加工及廢水之生物處理與土壤及地下水污染整治等。鑑於傳統微生物的鑑定,多採用分離培養鑑定,須長時間培養且環境中可培養之微生物有限;爾後發展之分子生物技術,如聚合酶連鎖反應(PCR);全細胞式定量法,如螢光原位雜交法(FISH),但仍然費時,且無法獲得全部資料。近年,新發展而成之技術,如流式細胞儀與生物晶片等,可快速鑑定微生物,但成本相對高昂,且無法同時鑑定多種微生物(n≧20)。為尋求更佳的微生物鑑定方法,本研究利用微機電工程之光微影製程,開發微流體生物晶片。本研究所開發之微流體生物晶片之微通道寬度與高度均在60 μm以下,利用微水珠封裝螢光分子染色之細菌,藉由雷射光激發及高靈敏度光學感應器擷取螢光訊號,再以類比轉數位介面卡及LabVIEWTM電腦軟體進行資料擷取統計與分析,期能達到快速鑑定與定量微生物之目的。

There is an unsolved problem in environmental ecology and environmental biology, i.e., a way to answer "who are they?", "how many are they of each species?" and "what are they doing?" for all the bacteria in an environmental sample. Those information can not only benefit basic research in environmental ecology and environmental microbiology but also assist other studies relating to microorganisms like biosynthesis, medicine, identity of pathogens, wastewater treatment, and soil and groundwater remediation.
Traditional way to identify microorganism is culturing methods, which are time consuming and unable to provide information on viable but not culturable bacteria. Thus, molecular biology methods have been developed such as polymerase chain reaction (PCR), fluorescence in situ hybridization (FISH), and so on. However, molecular biological methods are still time consuming and not of them is a portable technology. Recently, there are several kinds of new rapid technology being developed like flow cytometry, gene chip, and microarray. These technologies can identify microorganism in a fairly short period of time with high cost and still cannot answer the above research question.
Looking for much better answer for rapidly identifying microorganism, enumerating each species, and defining functional roles of each one of them, we employ photolithography, soft lithography, optical detection, and rapid data acquisition to develop micro-fluidic platform called Flow-Omics. Droplets which diameters are from 10 μm to 15 μm are formed in a specially-designed microfluidic chip. After labeling fluorescence molecules on each cell which has been packed in droplet, the intensity of fluorescence was detected and data are recorded by a data acquisition software, LabVIEWTM.
其他識別: U0005-2408201208133900
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