單一細胞操控、培養與即時觀測之研究

Abstract

細胞為生命之最小單位，個體的特殊性皆來自細胞的影響。幹細胞為身體各組織的原始細胞，擁有分化為其他不同細胞的能力，究竟是何種反應發號施令仍舊是個謎題，若能對單一幹細胞進行研究觀察，將有助於了解其中奧秘，因而科學家推動了單一細胞的研究。微機電系統技術可製作多種與生物分子尺寸相符的微小元件，有助於操控單一細胞，故結合微機電技術與生物醫學將利於單一細胞之研究。 本研究目的於建立一套單一細胞操控、培養與觀測之系統，研究方法可分為細胞分離與運送、培養與觀測分析。在細胞分離與運送方面，透過微機電製程輔助，對單一細胞進行操控。在細胞培養上，使用微型細胞培養室，將細胞培養於微量井晶片上，此晶片主要為提供單一細胞獨立成長環境，並利用晶片上的結構可對細胞進行培養液之替換或注入不同化學物質以刺激細胞，觀察細胞成長反應之現象。觀測系統則以全反射式螢光顯微鏡與CytovivaTM150等之顯微系統為主，即時觀測取得生物影像，並透過電腦軟體進行影像分析。 整合以上技術後，即需要生物知識來協助實驗研究，以正確解讀生物訊息，如細胞形變、蛋白質之運作等，進一步對單一細胞研究作更深入的探討。

Cell is the smallest, biologically functional system unit in lives, and every feature in organisms is constituted by cells. The most original cells in humans are stem cells which possess the remarkable potential to be developed into various cell types in body. The mechanism remains unknown such that a study of single stem cell is becoming a highly potential approach to unveil its activities. By taking the advantages in MEMS technique of comparable size to a cell, the manufacturing, manipulation and investigation for a single cell have been made in this work. The objective of the study is to develop a microsystem of single cell for manipulation, culturing, and real-time investigation. In manipulation for a single cell, a micro-pipette and micro-well device has been micromanufactured in order for isolation and transportation of a single cell. In cell culturing, a single cell is trapped and cultivated within a micro-well chip inside the micro-incubation culture system. The chip provides an isolated environment for a single cell in which the device is allowed to be injected with chemicals and biomarkers to stimulate the single cell. Moreover, in cell observation, the real-time investigation has been made simultaneously by both the total internal reflection fluorescence microscopy and CytovivaTM150. The images can be obtained and analyzed with the image processing software such that a great deal of bio-information for a single cell can be achieved. With such a system, the single cell morphology and protein-protein interaction can then be potentially realized. The well-integarted cell microsystem has been made for potential study for a single stem cell or its associated interaction, differentiation and communication to specific cells.