【作者】 王伟；

【导师】 南蓬； 罗源；

【作者基本信息】 复旦大学 ， 生物工程（专业学位）， 2013， 硕士

【摘要】 目的：石蜡切片是一种传统的切片方式,它主要用于镜下观察细胞形态,广泛应用于病理学和法医学领域。石蜡切片制作时还存在一些不足,如切片操作时间过长、固定液的效果不佳,封片溶液毒性过大等问题。皮肤组织由于结构复杂,在制片过程中具有一定的难度。本研究针对皮肤组织结构和成分的特点,对皮肤石蜡切片中固定液的选择、透明和封片溶剂毒性及效果进行比较,探讨不同方法中脱水,透明,浸蜡等程序设定的优劣,旨在找到适合于皮肤组织的快速,低毒的石蜡切片新方法。方法：选取2012年3月至2012年12月送检的成人包皮环切术所切下包皮组织样品25例,将每例样品分为10份,设置两组,传统方法组和新型方法组各为125份样品。新型方法组使用改进的脱水、透明、浸蜡、脱蜡、切片和封片方法进行制片。同时,将新型方法中的20个石蜡块样本放置室温1个月后再次进行切片、染色、封片。观察比较与1个月前新型方法切片得到的样本有无差异。计算传统切片法和新方法各步骤及总过程所用时间,比较甲基环己烷和二甲苯在透明过程和切片染色的效果,根据常规石蜡包埋-HE染色切片质量基本标准对两种石蜡切片方法进行质量评价。结果传统方法和新型方法制片都需经过12步来完成,新型方法组制片总过程所耗总时间(约7hrs)明显短于传统方法组(约83hrs)。其中新型方法组的脱水(45min),透明(20min),透蜡(21min),切片(10min),脱蜡(38min),封片(1h)都短于传统方法组的相对应步骤,脱水(5hrs),透明(1h),透蜡(70mmin),切片(30min),脱蜡(48min),封片(48hrs15min)。新型方法组的优级率与优良率明显高于传统方法组,且差异有统计学意义(p<0.05)；新型方法组切片质量总评分明显高于传统方法组,且差异有统计学意义(p<O.05)；其中新型方法组的组织切片完整性,组织切片厚度,切片有无刀痕,切片平整性,切片和载玻片间有无气泡,切片是否松散等项目评分明显高于传统方法组,且差异有统计学意义(p<0.05)；新型方法所取得的石蜡块存放一个月后与新鲜制备的石蜡切片并无明显差异。且传统方法制备的蜡块与新方法制备的蜡块共同存放5个月后也未表现出明显差异。更多还原

【Abstract】 Objective:Paraffin sections is a traditional way of slicing, it is mainly used for microscopic observation cell morphology, widely used in the field of pathology and forensic medicine. The paraffin sections also have some disadvantage, such as the selection of the fixative, the toxicity mounting and transparent reagents. Skin tissue have a very complex structure, the producer has a certain degree of difficulty. This study will focus on the characteristics of the structure and composition of the skin tissue, skin paraffin sections fixative choice, transparency and comparison of mounting solvent toxicity and effect, to discuss different methods of dehydrated, transparent dip wax program set to explore, and to find suitable for skin tissue fast, low toxicity paraffin section method.Method:25 adult foreskin samples were selected from the medical circumcision from March 2012 to December 2012. Each sample was divided into 10 small samples, and set two groups. The traditional method and the new method have 125 samples, respectively. The new method group was using a new type of dehydration, transparent, wax dipped, dewaxing, slided, mounting methods for production.. At the same time,20 embedded samples of new method were placed at room temperature for 1 month and then did sectioning, staining and mounting again. Compare with the differences between the traditional slicing method and the new method for process timing, methylcyclohexane and xylene effect in a transparent processing. According to the quality standards of paraffin-embedded sections, we evaluated the two methods.Result:Traditional methods and new methods both need 12 steps to complete the total process。The new method group production’s total time is about 7 hrs.The traditional methods group production’s total time is about 83hrs.The new method group was significantly shorter than the traditional method group.And the process of new method is dehydration (45min), transparent (20min), through the wax (21min), the slide (10min), dewaxing (38min), finalizing (1h). Meanwhile the traditional method group’s process is dehydration (5h), transparent (1H), transparent wax (70min), slide (30min) dewaxing (48min), and mounting (48hrs15min). Rate and excellent rate of the new method group level is significantly higher than the traditional methods group,and the difference was statistically significant (p<0.05). Slide quality of the new method group total score was significantly higher than the traditional method, and the difference was statistically significant (p<0.05). The integrity of the tissue sections; the thickness of tissue section; the slide marks on tissue section; the slided smoothness; The slicing and slides with or without bubbles of tissue section; whether tissue section loose items. All those technical keys above for new methods sample rated significantly higher than the traditional method group, and the difference was statistically significant (p<0.05).And 1 month delayed new method slides have no significant difference with freshly prepared new method slides.In the end traditional method prepared paraffin section and new method prepared paraffin section didn’t show any significant difference after 5 months storage in room temperature.更多还原