本文選題：粘附劑 + 固定液�。� 參考：《延邊大學(xué)》2017年碩士論文

【摘要】：冰凍切片技術(shù)是一種常用的獸醫(yī)病理學(xué)檢驗(yàn)技術(shù),主要用于獸醫(yī)臨床快速診斷、脂肪染色及組織內(nèi)抗原抗體反應(yīng)或酶的檢測。但獸醫(yī)臨床病理學(xué)診斷中,常常出現(xiàn)由于冰凍切片質(zhì)量不佳而判斷失誤的情況。本試驗(yàn)為提高冰凍切片質(zhì)量,以動(dòng)物的心臟、肝臟、脾臟、肺臟、腎臟及肌肉等為試驗(yàn)材料,利用常規(guī)組織學(xué)和冰凍切片技術(shù),進(jìn)行不同粘附劑、固定液及防冰晶法對冰凍切片質(zhì)量影響的研究。結(jié)果表明:1、不同粘附劑對固定組織冰凍切片質(zhì)量的影響。經(jīng)蛋白甘油、多聚賴氨酸、明膠硫酸鉻鉀三種粘附劑處理過的載玻片及從試劑公司購買的載玻片防脫片效果存在明顯差異。經(jīng)多聚賴氨酸處理過的載玻片的脫片率為0%,防脫片效果最佳,其次是明膠硫酸鉻鉀(脫片率為5%),而經(jīng)蛋白甘油處理過的載玻片和從試劑公司購買的載玻片脫片率達(dá)66%和92%,防脫片效果較差。2、不同固定液對新鮮組織冰凍切片質(zhì)量的影響。六種固定液短時(shí)間固定對新鮮組織冰凍切片的制片效果及H.E染色效果的影響各不相同。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、中性福爾馬林和AF固定液的效果差。3、不同防冰晶法對新鮮組織冰凍切片質(zhì)量的影響。不同防冰晶法均有去冰晶作用,但消除程度存在差異,其中液氮直接冷法和異戊烷處理25 s的異戊烷-液氮法效果最好,而蔗糖梯度脫水法和異戊烷處理20 s、30 s的異戊烷-液氮法效果均差。由此可知,制作一張高質(zhì)量的冰凍切片,應(yīng)采用以下方法:固定組織冰凍切片時(shí),采用多聚賴氨酸處理過的載玻片;制作新鮮組織冰凍切片時(shí),采用乙醚酒精1:1固定液固定及液氮直接冷卻法或異戊烷處理25 s的異戊烷-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.
【學(xué)位授予單位】：延邊大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S854.43

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