A Low Toxicity Method For Skin Tissue Paraffin Section

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.