冷冻切片固定质量影响因素的探讨

Discussion on the Factors Influencing the Fixation Quality of Frozen Section

目的探讨影响冰冻切片固定质量的因素。方法收集2016年1—12月厦门大学附属第一医院病理科的80例冷冻切片标本,每例标本连续切片7片,随机分为7组,A组电吹风快速吹干切片,B组立即放入95%乙醇固定1 min,C组立即放入10%中性福尔马林固定1min,D组立即放入丙酮固定1 min,E组立即放入AAF液固定1 min,M组立即放入AAF液固定15 s,N组立即放入AAF液固定30 s。固定后染苏木素、水洗、分化、返蓝、染伊红、脱水、透明、封固等,比较各组的脱片情况、染色结果。结果 A组未发现脱片现象,B、C、D、E组出现脱片的数量分别为15片、12片、24片和13片。将B组未脱片的65例编成F组,将C组未脱片的68例编为G组,将D组未脱片的56例编为H组,将E组未脱片的67例编为I组,比较A、F、G、H、I五组染色情况,I组组织清晰的切片比例明显高于A、F、G和H组(P均<0.05);在细胞皱缩度方面,与I组比较,F组、H组细胞均出现明显皱缩现象(P均<0.01);I组的细胞核染色鲜艳度和核浆对比度均明显优于其他四组(P均<0.05)。比较M、N和I组的染色情况,染色效果从优到劣依次为I组→N组→M组。结论 AAF是一种较为理想的冰冻切片固定液;影响冰冻切片固定质量的因素包括固定液的种类、冷冻组织的类别、固定时间的长短等。

Objective To investigate the factors affecting the fixation quality of frozen section. Methods 80 cases of frozen sections from the First Affiliated Hospital of Xiamen University were collected from January to December 2016. Each specimen was serially sectioned into 7 slices, randomly divided into 7 groups： A group, quickly drying slice with hair dryer. B group, immediately placed in 95% ethanol to fix 1 min. C group, immediately placed in 10% neutral formalin to fx 1 min. D group, immediately placed in acetone to fx 1 min. E group, immediately placed in the AAF liquid to fx 1 min. M group, immediately placed in the AAF liquid to fx 15 seconds. N group, immediately placed in AAF liquid to fx 30 seconds. After fixation, the process of dyeing hematoxylin, washing, differentiation, returning blue, dyeing eosin, dehydration, transparency, sealing and so on, were carried out according with the unifed procedure. The results of the unfalling and staining of each group were compared. Results No stripping was found in the A group. The number of stripping in B, C, D and E group was respectively 15, 12, 24 and 13. 65 cases of no stripping in B group were incorporated into F group. 68 cases of no stripping in C group were incorporated into G group. 56 cases of no stripping in D group were incorporated into H group. 67 cases of nostripping in E group were incorporated into I group. The staining of fve groups of A, F, G, H and I were compared. The number of clear tissue slices in I group was signifcantly higher than that of A, F, G and H group （P 〈 0.05）; In the aspect of cell shrinkage, compared with I group, the cells in F group and H group showed obvious shrinkage phenomenon （P 〈 0.01）; The nuclear staining brightness and contrast caryoplasm in I group were signifcantly better than the other four groups （P 〈 0.05）. The staining of M, N and I group was compared, the dyeing effect from priority to inferiority were I group, N group and M group. Conclusion AAF is an ideal fxation fuid for frozen section. The factors affecting the quality of frozen section include the types of the fxation liquid, the types of frozen tissue, the length of the fxation and so on.