Development of the Automatically Celluar Metabolic Analyzer with Ablility of Simultaneous Measurement of Three Metabolic Products

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Project title

Code/計畫編號

NSC102-2221-E019-015-MY2

Translated Name/計畫中文名

可同步偵測三種細胞代謝指標之自動化細胞代謝感測裝置雛形機開發

Project Coordinator/計畫主持人

Shih-Hao Huang

Funding Organization/主管機關

National Science and Technology Council

Department/Unit

Department of Mechanical and Mechatronic Engineering

Website

https://www.grb.gov.tw/search/planDetail?id=8123971

Year

2014

Start date/計畫起

01-08-2014

Expected Completion/計畫迄

01-07-2015

Bugetid/研究經費

827千元

ResearchField/研究領域

醫學工程

"商業機台「海馬生物能量測定儀(Seahorse Bioscience, USA, 2009)」是第一個使用24/96 孔盤為平臺，可即時同步偵測細胞有氧呼吸以及糖酵解作用的自動化機台，能快速篩選出具開發潛力之藥物及藥物毒性評估，然而，目前售價為10-20 萬美元與使用24/96 微孔盤，限制其普及性與應用性。本計畫預計以2 年時間開發具有多重藥物注入與同步偵測三種細胞代謝指標(OCR, OCAR, CDPR)之自動化細胞代謝感測裝置雛形機，可即時同步偵測有氧呼吸下的耗氧率O2(Oxygen Consumption Rate, OCR)、以及糖酵解作用下細胞外酸化率H+ (Extracelluar Acidification Rate, OCAR)、CO2 產率(Carbon Dioxide Production Rate, CDPR)，相較於商用機台，具有低售價、低細胞消耗量、可控制細胞培養微環境與多細胞共培養、與分析多種待測藥物…等優點。此裝置主要分三個部份：(1)細胞培養微陣列晶片：在玻璃基材上製作出微陣列孔洞晶片，將感測磷光薄膜製作於孔洞底部，並將細胞培養貼附於微陣列孔洞內，可進行長時間細胞培養。(2)氣動密閉作動裝置：具有可氣壓上下致動的閥座密封上述細胞培養微陣列晶片中的微陣列孔洞，具有藥物注入導引微流道與流體驅動裝置，可依據需求注入多種藥物。(3)磷光相位差光學檢測系統：可控制激發光的光源與投影圖像，激發上細胞培養微陣列晶片中的微陣列孔洞內的感測磷光薄膜，並透過磷光相位差計算處理，藉此得知孔洞內的細胞三種細胞代謝指標。透過三種細胞代謝指標(OCR, OCAR, CDPR)參數的檢測，以及具有藥物注入導引流體驅動裝置，可依據需求注入多種藥物，以便研究不同藥物對細胞新陳代謝的影響，快速解析待測細胞的基礎代謝率、ATP 轉換、氫離子滲漏（膜的完整性）、極限呼吸率。透過此開發機台的協助，可廣泛應用於癌症、老化、心血管、細胞生理、線粒體疾病、神經退化、肥胖症、糖尿病、與各種疾病機制、藥物開發與篩選的研究上。" "XF Extracellular Flux Analyzers (Seahorse Bioscience, USA, 2009) available in 24-well and 96-well formats are the industry standard for measuring cellular bioenergetics, simultaneously measuring the two major energy producing pathways of the cell – mitochondrial respiration and glycolysis - in a microplate, in real-time. The local distributor in Taiwan is Cell-bio biotechnology Co., Ltd. However, the high cost and the usage of 24/96 microplates limit its applications and popularization. In this project, we plan to spend two-year efforts for development of the automatically celluar metabolic analyzer with ability of simultaneous measurement of three metabolic products, i.e., oxygen consumption rate (OCR), extracelluar acidification rate, OCAR), carbon dioxide production Rate (CDPR). The device consists of three main components including: (1) A microwell array for single cell culture. Cells were seeded onto the sensitive dyes inside a microwell. (2) Pneumatically-driven lid to hermetically seal the microwell. This component also contains microfluidic channels for automatic drug injection. (3) A optical measuring system for measurements of cellular OCR, OCAR, and CDPR based on phase-based lifetime detection. This system includes a digital micromirror device (DMD) to project an illumination patterns, which excites the sensitive dyes for measurement of OCR, OCAR, and CDPR. Through our proposed device, scientists can have new insights into obesity, diabetes, cancer, cardiovascular and neurodegenerative diseases."

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Development of the Automatically Celluar Metabolic Analyzer with Ablility of Simultaneous Measurement of Three Metabolic Products

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