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一种用于组织切片冷冻保存的新型简化超快速冷冻技术。

A novel simplified ultra-rapid freezing technique for cryopreservation of tissue slices.

作者信息

Rypka Miroslav, Cervenková Katerina, Uherková Lenka, Poczatková Hana, Florschutz Anthony V, Veselý Jaroslav

机构信息

Department of Pathological Physiology, Faculty of Medicine, Palacký University, Olomouc, Czech Republic.

出版信息

Cryobiology. 2006 Apr;52(2):193-9. doi: 10.1016/j.cryobiol.2005.10.012. Epub 2005 Dec 9.

Abstract

Cryopreservation offers the potential to maximize the use and availability of biological materials that have a limited supply. This study demonstrates an enhanced technique for the parallel cryopreservation of a series of liver tissue slices using a tray modeled from aluminium foil and low concentrations of a cryoprotectant. Cooling and warming rates of approximately 2000 and 3900 degrees C min(-1), respectively, were achieved as the thermal capacity of the foil-tray was significantly reduced compared to the aluminium sandwich device introduced by Day et al. [S.H. Day, D.A. Nicoll-Griffith, J.M. Silva, Cryopreservation of rat and human liver slices by rapid freezing, Cryobiology 38 (1999) 154-159]. Additionally, the two critical steps involved in the sandwich approach, i.e., clamping the plates and complete filling of the entire space between the plates with liquid, can be omitted using the foil tray. The viability of the slices was verified by measuring tetrazolium salt reduction capacity, cytosolic enzyme lactate dehydrogenase leakage, and ethoxycoumarin metabolism.

摘要

冷冻保存为最大限度地利用供应有限的生物材料提供了可能。本研究展示了一种改进技术,可使用由铝箔制成的托盘和低浓度冷冻保护剂对一系列肝组织切片进行并行冷冻保存。由于与Day等人介绍的铝制夹层装置相比,箔盘的热容量显著降低,分别实现了约2000℃/分钟和3900℃/分钟的冷却和升温速率。[S.H. Day, D.A. Nicoll-Griffith, J.M. Silva, Cryopreservation of rat and human liver slices by rapid freezing, Cryobiology 38 (1999) 154 - 159]。此外,使用箔盘可以省略夹层方法中涉及的两个关键步骤,即夹紧板和用液体完全填充板之间的整个空间。通过测量四唑盐还原能力、胞质酶乳酸脱氢酶泄漏和乙氧基香豆素代谢来验证切片的活力。

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