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題 名 | 應用微擴散室系統測定九孔卵之滲透壓特性 |
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作 者 | 林達德; | 書刊名 | 國立臺灣大學農學院研究報告 |
卷 期 | 32:1 1992.03[民81.03] |
頁 次 | 頁31-48 |
分類號 | 439.61 |
關鍵詞 | 九孔卵; 微擴散室系統; 滲透壓; |
語 文 | 中文(Chinese) |
中文摘要 | 生物細胞在低溫冷凍或解凍過程中,由於細胞膜外界形成冰晶而造成細 胞內外滲透壓不平衡之現象,因此以水滲透作用來達成細胞內外的滲透壓平衡, 平衡過程之快慢則視細胞之滲透壓特性而定。故細胞滲透壓特性之測量將有助於 細胞低溫保藏程序中最佳冷凍速率之判斷,對於細胞於低溫保藏過程中死亡的機 制方可由數學模式經電腦模擬而更深入瞭解。另外,對於目前常應用於低溫保存 之急速冷凍玻化法中,冷凍保護劑添加移除程序的改善或新程序的建立亦為重要 之資訊。傳統測定細胞滲透壓特性的方法多為應用光線吸收或散射的原理,這些 方法通常需要較大,的測試細胞,且由於所測得之細胞滲透壓特性為整個試樣的 平均值,單一細胞之滲透壓特性無法測得。針對傳統滲透壓測量法之缺點,應用 滲析薄膜所設計的微拭散室系統可以輕易的在顯微鏡下測得單一細胞的動態及 靜態滲透壓特性。本研究之目的在應用新近發展的微擴散室技術建立一套完整的 單細胞滲透壓特性測量系統,同時川九孔卵為對象完成其滲透壓特性之測量與分 析。應用微擴散室系統所測得九孔卵之平衡體積比與滲透壓濃度成線性反比關 係。迴歸分析結果求得九孔卵之靜態透滲透皮特性可以Boyle-Van'tHoff關係表示 為FV=0.454/C+0.540,其中FV為平衡體積比,而c為滲透壓濃度。式中之0.540為 縱軸截距,此值顯示九孔卵中參與滲透壓作用之水份並不高。動態實驗中所求得 體積比對時間之數據,以數學模式利用非線性迴歸方法分析九孔卵之水滲透係數 Lp,求得九孔卵在溫度20℃時之Lp值為6.36±2.30x1O���沸/(s-atm)。再於不同溫 度下進行實驗而進一步分析溫度對於Lp之影響,不同溫度下所求得之Lp值繪於 Arrhenius圖中呈線性關係,而依此關係計算出水滲透係數Lp之活化能△E為11.6 ±0.6kcal/mole。 |
英文摘要 | The use of the recently developed micro-diffusion chambertechnique has facilitated the determination of the osmometriccharacteristics of single biological cell. The determined osmometricparameters such as hydraulic conductivity and solute permeability canbe used in mathematical models for the prediction of osmometricbehavior of cells subject to various cryopreservation protocols. Withthese predictions, the mechanisms which cause the decrease of cellsurvival during freezing can be better understood. Traditionalcryopreservation protocols can thus be improved or new protocols canbe established to increase the survival of cryopreserved cells. The goalof this research is to establish a measuring system for thedetermination of the osmometric characteristics of single biologicalcell based on the micro-diffusion chamber technique. Eggs of smallabalone were selected as test specimen, and their osmometriccharacteristics were determined and analyzed using the micro-diffusionchamber system. The static osmometric characteristics of smallabalone eggs was determined and analyzed. In the Boyle-Van't Hoffplot each datum of individual eggs was regressed and the Boyle-Van'tHoff relation was determined to be FV=0.454/C+0.540, where FV is theequilibrium fractional volume and C is the osmolality of the suspendingsolution. The value of intercept, 0.540, indicates the small abaloneeggs contain high percentage of osmotically inactive materials. Thedata from dynamic measurement were fitted to a mathematical modelfor the determination of hydraulic conductivity Lp using the non-linearregression method. Estimates of Lp among individual eggs were highlyvariable. The estimated Lp at 20℃. were 6.36±2.30 x 10���沸/(s-atm). Inthe Arrhenius plot a linear relationship was regressed which yieldedthe activation energy (AE) of Lp to be 11.6± 0.6 kcal/mole. |
本系統中英文摘要資訊取自各篇刊載內容。