固定液温度对冰冻切片质量的影响

Effect of fixation fluid temperature on the quality of frozen sections

目的探讨固定液温度对冰冻切片质量的影响。方法选取南方医科大学南方医院冰冻新鲜肺组织20例、乳腺组织20例、肠组织20例、甲状腺组织20例和脑组织20例,每个病例取一块组织,每块组织连续切片4片,分成A组、B组、C组、D组,每组100片。A组采用室温AA固定液中固定1 min;B组采用低温恒冷AA固定液中固定1 min;C组采用低温恒冷AA固定液中固定1 min,然后热风固定10 s;D组采用低温恒冷固定液中固定15 min;常规苏木精-伊红染色法(HE染色)进行HE切片染色,观察四组的细胞形态差异、细胞核和胞浆的染色效果。结果A、C组冰冻切片组织细胞轻微收缩,HE染色均匀,细胞核染色深蓝色,核浆对比度较好;B组冰冻切片细胞轻微肿胀,切面易发生裂隙,HE染色不均匀,细胞核染色灰白灰蓝色,核浆对比度较差;D组冰冻切片组织细胞局部轻微肿胀,HE染色局部不均匀,细胞核染色深蓝色或灰蓝色,核浆对比度较差。A组固定液染色综合评分为(93.99±0.69)分,B组固定液染色综合评分为(85.95±1.16)分,C组固定液染色综合评分为(93.61±0.62)分,D组固定液染色综合评分为(91.85±0.57)分,四组固定液染色综合评分比较,差异有统计学意义(P<0.05)。结论组织冰冻切片经室温AA固定液固定后,细胞形态自然,HE染色细胞核染色鲜艳,核浆对比度较好。

Objective To discuss the effect of fixation fluid temperature on the quality of frozen sections.Methods 20 cases of frozen fresh lung tissue,20 cases of breast tissue,20 cases of intestinal tissue,20 cases of liver tissue,20 cases of thyroid tissue and 20 cases of brain tissue in Nanfang Hospital of Southern Medical University were selected.1 piece of tissue was taken from each case,and each piece of tissue was cut into 4 pieces,which were divided into group A,group B,group C and group D,with 100 pieces in each group.Group A was fixed in room temperature AA fixation fluid for 1 min.Group B was fixed in low temperature and constant cooling AA fixation fluid for 1 min.Group C was fixed in low temperature and constant cold AA fixation fluid for 1 min,followed by hot air fixation for 10 s.Group D was fixed in low temperature and constant cooling fixation fluid for 15 min.Hematoxylin eosin staining(HE staining)was used to observe the cell morphology,nuclear and cytoplasmic staining effect of the four groups.Results In group A and C,the frozen section cells contracted slightly,the HE staining was evenly,the nuclear staining was dark blue,and the contrast of nuclear plasma was good.In group B,the cells in frozen section were slightly swollen,the section was easy to crack,the HE staining was uneven,the nucleus staining was gray and gray blue,and the nucleocytoplasmic contrast was poor.In group D,the frozen section tissue cells were slightly swollen,the HE staining was uneven,and the nuclear staining was dark blue or gray blue,and the nucleocytoplasmic contrast was poor.The comprehensive score of fixation fluid staining of group A was(93.99±0.69)points,which was(85.95±1.16)points in group B,(93.61±0.62)points in group C and(91.85±0.57)points in group D.There was statistically significant difference in comprehensive score of fixation fluid staining among the four groups(P<0.05).Conclusion After frozen tissue sections are fixed with room temperature AA fixation fluid,the morphology of the cells is natural,the nucleus stain with HE is stained brightly,and the nucleocytoplasmic contrast is good.