不同粘附剂、固定液及防冰晶法对冰冻切片质量的影响

发布时间：2018-06-21 23:33

本文选题：粘附剂 + 固定液　；参考：《延边大学》2017年硕士论文

【摘要】：冰冻切片技术是一种常用的兽医病理学检验技术,主要用于兽医临床快速诊断、脂肪染色及组织内抗原抗体反应或酶的检测。但兽医临床病理学诊断中,常常出现由于冰冻切片质量不佳而判断失误的情况。本试验为提高冰冻切片质量,以动物的心脏、肝脏、脾脏、肺脏、肾脏及肌肉等为试验材料,利用常规组织学和冰冻切片技术,进行不同粘附剂、固定液及防冰晶法对冰冻切片质量影响的研究。结果表明:1、不同粘附剂对固定组织冰冻切片质量的影响。经蛋白甘油、多聚赖氨酸、明胶硫酸铬钾三种粘附剂处理过的载玻片及从试剂公司购买的载玻片防脱片效果存在明显差异。经多聚赖氨酸处理过的载玻片的脱片率为0%,防脱片效果最佳,其次是明胶硫酸铬钾(脱片率为5%),而经蛋白甘油处理过的载玻片和从试剂公司购买的载玻片脱片率达66%和92%,防脱片效果较差。2、不同固定液对新鲜组织冰冻切片质量的影响。六种固定液短时间固定对新鲜组织冰冻切片的制片效果及H.E染色效果的影响各不相同。其中乙醚酒精1:1固定效果效果最好,其次是AAF固定液、4%多聚甲醛,而95%乙醇、中性福尔马林和AF固定液的效果差。3、不同防冰晶法对新鲜组织冰冻切片质量的影响。不同防冰晶法均有去冰晶作用,但消除程度存在差异,其中液氮直接冷法和异戊烷处理25 s的异戊烷-液氮法效果最好,而蔗糖梯度脱水法和异戊烷处理20 s、30 s的异戊烷-液氮法效果均差。由此可知,制作一张高质量的冰冻切片,应采用以下方法:固定组织冰冻切片时,采用多聚赖氨酸处理过的载玻片;制作新鲜组织冰冻切片时,采用乙醚酒精1:1固定液固定及液氮直接冷却法或异戊烷处理25 s的异戊烷-液氮法防冰晶。
[Abstract]:Frozen section technique is a commonly used technique for veterinary pathological examination, which is mainly used for rapid diagnosis of veterinary clinic, fat staining and detection of antigen-antibody reaction or enzyme in tissues. However, in veterinary clinicopathological diagnosis, errors often occur due to poor quality of frozen sections. In order to improve the quality of frozen sections, the animals' hearts, liver, spleen, lungs, kidneys and muscles were used as experimental materials. Study on the effect of immobilization solution and antiglacial method on the quality of frozen sections. The results showed that the effect of different adhesives on the quality of frozen sections of fixed tissues was observed. The anti-stripping effect of the slide treated with three adhesives, protein glycerin, poly-lysine and glutin chromium sulfate potassium sulfate, was significantly different from that of the glass slide purchased from the reagent company. The removal rate of the glass slide treated with poly-lysine is 0, and the anti-stripping effect is the best. The second was gelatin chromium sulfate potassium sulfate (5% stripping rate), while 66% and 92% were obtained for glass slides treated with proteoglycerin and those purchased from reagent company. The anti-stripping effect was not good. The effects of different fixations on the quality of frozen sections of fresh tissues were studied. The effects of short time fixation of six kinds of fixatives on the preparation of frozen sections of fresh tissue and the effect of e staining were different. The effect of ether alcohol fixation at 1:1 was the best, followed by AAF fixative solution (4% paraformaldehyde), while the effect of 95% ethanol, neutral formalin and AF fixation solution was poor. The effect of different antiglacial methods on the quality of frozen sections of fresh tissue was studied. Different antiglacial methods had deglacial effect, but the degree of elimination was different. The isopentane liquid nitrogen method with direct cooling of liquid nitrogen and isopentane for 25 s had the best effect. However, the effect of sucrose gradient dehydration and isopentane treatment for 20 s ~ 30 s by isopentane-liquid nitrogen method was poor. It can be concluded that the following methods should be used to make a high quality frozen slice: when fixing frozen tissue sections, using polylysine treated slides; when making frozen slices of fresh tissue, Isopentane-liquid nitrogen method for 25 s was used to immobilize ice crystals with ether alcohol 1:1 stationary solution and direct cooling of liquid nitrogen or isoamyl alkane.
【学位授予单位】：延边大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S854.43

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