近年來光學技術已廣泛應用在醫學診斷。本質性光訊號是量測組織細胞光學特性能力變化的重要方法，用以觀察組織細胞在各種之生化調控條件下的光反應情形，做為推測組織細胞生理狀態的參考依據。此外，由持續的細胞膜重組行為所造成的細胞微動現象，亦是評估細胞活動力的重要指標。對於中風救治，適當的治療介入時機是能否救治瀕臨受損之神經細胞的最為重要關鍵；細胞在中風環境下隨時間演進的生理參數變化，是作為中風治療時機與治療方法的評斷依據。一般常見的中風實驗環境乃是抽離密閉式艙巢內氧氣達到缺氧狀態，此種作法限制了實驗取樣頻率。
本研究目的即在於運用光纖技術進行細胞生理狀態監控；連續、長時間偵測並評估培養於施以神經生長因子PC12細胞，在非封閉缺氧缺糖環境下，背向光散射訊號及細胞微動之光變化情形。實驗平台由本質光訊號量測、非封閉式室缺氧缺糖中風環境調控二項系統組成。以高低張溶液調控細胞組織特性，進行單光路、二光路兩類光纖探頭之本質光偵測效益評估。並以就地持續供應氮氣、缺氧缺糖溶液循環等兩種方式評估非封閉式缺氧艙巢的缺氧程度與穩定度。經由整合兩系統，測量PC12細胞在自正常培養環境到施以4小時缺氧缺糖環境，期間之本質光振幅，及低頻光亮變化。
本實驗結果顯示，本質光量變化呈現低張環境下，光量上升，反之則光量減少。以就地持續方式供應氮氣方式可提供最佳缺氧環境；此外，於缺氧艙巢外圍加以水槽之設計，能確保細胞培養環境不會因為大量氮氣流入帶走溶液內水分，而導致高張溶液環境。由光量之強度頻譜分布分析，頻譜強度隨施以缺氧缺糖環境時間的增加，而呈現逐漸衰減的趨勢。本實驗之獨特設計，提供以即時、連續及非化學標記的中風細胞生理狀態監控平台，致力於提供中風徵兆偵測與作為適時治療之參考依據。

Optics technologies have been widely applied in medical diagnosis and therapy. Intrinsic optical signal (IOS) is the important approach to observing the biologic tissue under varied physical or chemical conditions by measuring the properties of light scattering and absorption. Cellular micromotion is the integral behavior of continuous cell membrane remodeling which has been used as an indicator for evaluating the cell’s vitality and motility. Treatment of stroke at appropriate time is critical to rescue the nearly damaged cells within penumbral zone. Using micromotion detection to determine the interstate of cells when they are insulted with ischemic condition has been less investigated. Traditional hypoxia chamber is well-developed device for ischemia experiment but the enclosed chamber design limits frequent sampling.
The aims of thesis study are to apply the fiber optics technique to time-course monitor the optical response modulated by the varying micromotion of ischemic NGF-differentiated PC12 cells. Two types of fiber-optical probes were evaluated by sensing backscattering under varied osmotic conditions. The deoxygenated environment was created by using direct supply of nitrogen to culture medium or indirect approach of supplying the pre-prepared deoxygenated medium. The integration of a compact optical probe and a non-enclosed hypoxia chamber has been implemented and validated to continuously sense the backward optical scattering from cells’ membrane surface while they were kept in normal medium then underwent ischemic insult.
Our results indicated that the backscattering intensity was found to be inversely proportional to osmotic changes. The design of continuous supply of nitrogen into the water-trapped chamber not only can induce the most stable deoxygenated environment but also prevent the dehydration effect caused by nitrogen pumping. The quantitative varying spectrum slope showed that the cellular micromotion resulting in low frequency fluctuation would be with higher intensity while cells were healthy but gradually decline after they were injured by a period of ischemic insult. Our novel design makes it possible to proceed a label-free, real-time and continuous interstate monitoring for cells undergo ischemic insult.

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