本文關鍵詞： 蠕形螨 環(huán)境電子顯微鏡(ESEM) 超微結構 DNA提取　出處：《山東大學》2005年碩士論文　論文類型：學位論文

【摘要】：目的 應用環(huán)境掃描電子顯微鏡(environmental scanning electron microscope,ESEM)技術觀察人體蠕形螨的超微結構,確定其口器性質,尋找其與致病和分類有關的超微結構。用客觀證據(jù)說明蠕形螨是否致病,做出超微結構分類并對原形態(tài)學分類進行評價。 提取兩種人體蠕形螨基因組,為人體蠕形螨分子生物學研究鋪平道路。 方法 用刮片式取螨器從成人額部取螨,置于2%洗潔精溶液中,在解剖鏡直視下,用自制挑螨器分離清洗干凈。 將潔凈活螨按觀察部位的要求,在解剖鏡下擺上電鏡樣品托,立即掃描,拍攝照片。環(huán)境掃描電鏡工作參數(shù):工作距離(WD)7.5-9.3mm、高電壓(HV)20.0KV、高真空探頭(Det)GSED、束斑(Spot)3.0、溫度5℃、壓力0.5kpa、相對濕度60%。 將潔凈新鮮的毛囊蠕形螨(Demodex folliculorum, D. f. )、皮脂蠕形螨(Demodex brevis, D. b. )分別在解剖鏡直視下研磨粉碎。用改進的小型昆蟲基因組提取方法,分別提取兩種人體蠕形螨的基因組DNA,將提取物進行瓊脂糖凝膠電泳和紫外分光度計檢測。 結果 D. f. 的口器位于顎體腹面�？诳资莾善谙掳逯虚g的狹長縱向裂隙,有一劍峰狀的口針從裂隙中向前下方伸出,但未見牙齒。推斷其口器是一種特殊的刺吸式口器。D. b. 的口器與D. f. 相似,但尚未見口針伸出。 D. f. 須肢位于顎體兩側,分三節(jié),末節(jié)可向腹面做超過90°的運動。末端似釘鞋底狀,其上有7個須肢爪,刺狀。前三個呈三角形排列,后四個緊密排成兩排。D. b. 須肢較短,末節(jié)上只有5個須肢爪。 D. f. 口下板為梨形,外板較平;D. b. 口下板短而寬,呈寬橢圓形,外板唇狀。
[Abstract]:Objective to apply environmental scanning electron microscope (ESEM) to environmental scanning electron microscope. ESEM technique was used to observe the ultrastructure of human Demodex, to determine its oral structure, and to search for its ultrastructure related to pathogenicity and classification. Objective evidence was used to explain whether Demodex was pathogenic or not. Ultrastructural classification was made and the original morphological classification was evaluated. Two species of human Demodex genomes were extracted to pave the way for the molecular biology of human Demodex mites. Methods the mites were extracted from the frontal part of adult with scraper and placed in the solution of 2% detergent. The mites were separated and cleaned by self-made mite picker under anatomic microscope. The clean live mites were placed under anatomic microscope and scanned immediately according to the requirements of observation site. The working parameters of environmental scanning electron microscope were as follows: working distance was 7.5-9.3mm. High voltage HVV 20. 0 KV, high vacuum probe Dett GSED, beam spot 3. 0, temperature 5 鈩，

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