Autofluorescence and spectrum characteristics of Echinococcus granulosus culturing in vitro / 中国人兽共患病学报

ABSTRACT

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 .