方向式低溫顯微鏡系統之研製與應用

Implementation and Application of a Directional Cryomicroscope System

龍侃(Ken Lung) ； 林達德(Ta-Te Lin)

《農業機械學刊》 3卷4期 (1994/12) Pp. 15-24

https://doi.org/10.30062/JAM.199412.0002

摘要

低溫顯微鏡為觀察生物細胞在低溫狀態下所產生外觀變化的最主要儀器，本研究之主要目的為改良原有方向式低溫顯微鏡冷凍臺之設計，並以微電腦為基礎進行冷凍臺之溫度控制。所研製完成的方向式低溫顯微鏡系統主要包括冷凍臺座、熱源和熱淵及其溫度控制系統、樣本溫度讀取界面、定位機構與驅動電路及樣本溫度控制軟體等五個子系統。系統於研製完成後進行組合與性能測試，其可控制之溫度範圍為-80℃至100℃。在熱源和熱淵溫度設定為40℃與-80℃的條件下，控制恆溫之平均平方根誤差約為0.3℃，冷凍速率可達-60℃/min。所完成之低溫顯微鏡系統並以牡蠣卵與胚體為對象進行實驗，實驗結果顯示，隨著時間的增長，胚體與卵發生IIF的機率隨之增加，且隨著所設定的恆溫之下降，發生IIF的機率也隨之增加。胚體與卵之(上標 50)T(下标 IIF)分別為-21.0℃與-22.5℃左右，而牡蠣胚體之IIF機率由0增加到1的轉變溫度範圍較卵為寬廣。

關鍵字

低溫顯微鏡 ； 冷凍 ； 細胞內凍結 ； 牡蠣卵

並列摘要

Cryomicroscope is the most important instrument for the real-time observation of physical changes of biological cells at low temperature. The objectives of this research were to improve the original design of the directional cryomicro-scope and to utilize microcomputer for temperature control. The cryomicro-scope system was implemented and comprised five subsystems. They were: the cold stage base, the heat source and heat sink with independent temperature control systems, the sample temperature acquisition interface, the positioning mechanism and interface, and the sample temperature control software. The five subsystems were integrated and the performance of the cryomicroscope system was tested. The sample temperature could be controlled in the range between -80℃ and 100℃. When the temperatures of heat source and heat sink were set at 40℃ and -80℃, the root mean squared error of isothermal control was about 0.3℃. A maximum cooling rate of -60℃/min was obtained. The cryomicroscope system was also used to measure the IIF characteristics of oyster eggs and embryos. The IIF process of oyster eggs and embryos was found to be time and temperature dependent. The probability of IIF increased with time and lower temperature. The isotherm 50T(subscript IIF)of eggs and embryos were determined to be -21.0℃ and -22.5℃. The transition temperature zone of IIF probability from 0 to 1 appeared to be broader for embryos than that of eggs.

並列關鍵字

Cryomicroscope ； Freezing ； Intracellular Ice formation ； Oyster eggs