比较三种防冰晶法对切片保存效果的影响

目的 比较冰冻切片技术中几种防冰晶方法对切片保存效果的影响。方法 分别使用液氮骤冷组织、高渗透压脱水法、单纯OCT胶包埋等几种方法后行冰冻切片、HE染色后，分别于当天、1、3、6个月后比较其染色效果，选出最佳保存方法。结果 单纯OCT包埋法在1个月后染色明显变浅，冰晶数量最多，而在3、6个月后染色基本接近背底色，冰晶面积已占据近半个视野，组织结构破坏极其严重；高渗透压脱水法在1个月后染色深度及冰晶数量上无明显变化，但3、6个月后则明显变浅，冰晶数量明显增加；而低温骤冷法在染色后的几个月，染色深度及冰晶的数量均无明显的变化，结构基本维持染色当天的状态。结论 低温骤冷法是这三种防冰晶法中最利于保存切片的方法。

COMPARISON OF THE INFLUENCE OF ANTI-ICE CRYSTAL METHODS ON SECTION PRESERVATION

Objective To compare of the influence of three anti-ice crystal methods on section preservation.Methods The samples treated by liquid nitrogen rapid freeze method, high osmotic pressure dyring method, or simple OCT fixation method were frozen sectioned and HE stained. Then the signal strength of each sample was examined at three time points (the same day of staining, and 1,3 and 6 months after).Results The stain on samples treated with simple OCT fixation method started fading 1 month later, and many ice crystals could be seen; after three or six months the stain nearly ran out, the ice covered nearly half of the section, and tissue structure was seriously damaged. The samples treated with high osmotic pressure methods faded and formed ice slower than OCT fixation method treated ones. The tissue treated with liquid nitrogen rapid freeze method could maintain the color and effectively prevent ice formation even after 6 months. Conclusion The liquid nitrogen rapid freeze method is the best anti-ice crystal mation method for section preservation.