中文摘要：

目的研究细粒棘球蚴的自发荧光现象及共聚焦λ扫描特点,丰富细粒棘球蚴光谱学和生物学信息,为后期免疫荧光及医学光子学研究奠定基础。方法采集自然感染细粒棘球蚴的绵羊肝脏,在无菌条件下收集包囊内的原头蚴进行体外培养,亚甲基蓝染色确定原头蚴活力。激光扫描共聚焦显微镜下分别以405nm、488nm、514nm、561nm等不同激发光激发观察虫体自发荧光,并分别对这5种激发光激发的细粒棘球蚴做自发荧光λ扫描分析,同时采用全波长多功能酶标仪下分别以405nm、488nm、514nm、563nm、633nm等5种激发光激发细粒棘球蚴,绘制其自发荧光曲线。结果经亚甲基蓝染色测定原头蚴活力为100%。共聚焦显微镜观察不同激发光照射下细粒棘球蚴虫体能发出多种不同颜色的自发荧光。在相同的激发强度及探测器电压的情况下,以405nm激光激发的蓝色荧光效果最佳,虫体大体结构清晰;λ扫描分别以405nm、488nm、514nm、561nm、633nm等5种激发光激发样品,相应敏感的发射波长分别为490~520nm、520nm及580nm左右、580~600nm、610~630nm和650nm左右,其中405nm、488nm和561nm激发效果较好,514nm、633nm激发效果欠佳。全波长多功能酶标仪检测细粒棘球蚴做自发荧光曲线与共聚焦显微镜结果基本一致,但所测定的虫体自发荧光强度较共聚焦显微镜偏低。结论激光扫描共聚焦显微镜下可观察到细粒棘球蚴虫体自发荧光,其中以405nm激发的蓝色荧光最强;细粒棘球蚴虫体自发荧光光谱较宽,490~520nm、610~630nm,405nm、488nm和561nm为较适宜的激发波长。

英文摘要：

The objective of this study was to illustrate the autofluorescence phenomenon and scanning confocal λcharacteris‐tics of Echinococcus granulosus （Eg） cultured in vitro ,which can not only rich the spectroscopy and biological information of Eg ,but also lay a foundation for study the immunofluorescence and medical photonics .Protoscoleces （PSC） of Eg were aspi‐rated and pooled from sheep liver hydatid cysts collected from a slaughterhouse .The PSC viability was determined by Methyl‐ene blue staining .The autofluorescence of endocyst were observed through the confocal microscope excited respectively by 405 nm ,488 nm ,514 nm ,and 561 nm .The autofluorescence curve was measured by thermo Varioskan Flash respectively by 405 nm ,488 nm ,514 nm ,561 nm ,and 633 nm .Theλ scanning analysis was executed by confocal respectively by 405 nm ,488 nm ,514 nm ,561 nm ,and 633 nm to measure the autofluorescence curve .The Eg excited by different excitation light through confocal showed different colors autofluorescence .Under the same excitation intensity and detector voltage condition ,blue fluo‐rescence showed the best fluorescence effect and clearer structure which excited by 405 nm laser .Scanning respectively by five different excitation lights ,the corresponding sensitive emission wave lengths scope were 490‐520 nm ,around 520 nm and 580 nm ,580‐600 nm ,610‐630 nm ,and around 650 nm ,respectively .The 405 nm ,488 nm and 561 nm excitation effect were bet‐ter than that of the 514 nm and 633 nm .The autofluorescence curve detected by Microplate Reader were basically as same as that by confocal ,although the intensity detected by Microplate Reader were lower .The results indicated that the best excita‐tion light to detect the autofluorescence of Eg is 405 nm ,which showed blue fluorescence . The autofluorescence spectrum of Eg is wide ,mainly in 490‐520 nm and 610‐630 nm .The better excitation wave lengths are 405 nm ,488 nm and 561 nm .