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当前位置: 首页 > 文章 > 异硫氰酸荧光素标记鳗弧菌条件优化及应用条件对荧光信号的影响 水产学报 2014,38 (4) 132-139
Position: Home > Articles > Optimizing conditions of fluorescein isothiocyanate labeled Vibrio anguillarum and the effect of application condition on fluorescence signal Journal of Fisheries of China 2014,38 (4) 132-139

异硫氰酸荧光素标记鳗弧菌条件优化及应用条件对荧光信号的影响

作  者：
王锐;王秀华;王玉娟;韩雯;黄健
单  位：
中国海洋大学海洋生命学院;中国水产科学研究院黄海水产研究所
关键词：
鳗弧菌;异硫氰酸荧光素;荧光标记;标记条件
摘  要：
为了实现对目标细菌的有效示踪,实验以异硫氰酸荧光素(FITC)为标记物,分别设置FITC标记浓度0.2、2、20、50、100、200μg/mL,标记时间1、15、30、60、120、240 min,研究了FITC标记浓度与标记时间对鳗弧菌标记效果的影响,同时对在不同温度(25、4、-20、-70℃)及不同时间(24、48、96、192 h)存储条件下鳗弧菌标记荧光的稳定性进行了比较,对标记鳗弧菌经甲醛灭活后的荧光变化进行了观察,研究了不同光照条件(0、150、1 500、25 000 lx)对标记鳗弧菌的荧光衰减影响,以及在-20和-70℃条件下反复冻融对标记鳗弧菌荧光强度的影响。结果表明,FITC的最适标记浓度范围为20~100μg/mL,最适标记时间范围60~120 min,储藏在-20和-70℃条件下,标记后的鳗弧菌荧光信号比储藏在4和25℃条件下稳定,标记后的鳗弧菌经甲醛灭活后的荧光信号稳定性优于对照组,不同强度自然光照对标记鳗弧菌的荧光衰减效果不明显,标记细菌经3次冻融,发现在-70℃条件下冻存的细菌荧光信号稳定性保持良好。本研究结果为FITC用于标记海洋细菌提供了实用的技术参数。
译  名：
Optimizing conditions of fluorescein isothiocyanate labeled Vibrio anguillarum and the effect of application condition on fluorescence signal
作  者：
WANG Rui;WANG Xiuhua;WANG Yujuan;HAN Wen;HUANG Jie;College of Marine Life Science,Ocean University of China;Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences;
关键词：
Vibrio anguillarum;;fluorescein isothiocyanate;;fluorescence labeling;;labeling conditions
摘  要：
In order to track the target bacteria effectively,fluorescein isothiocyanate( FITC) was used as the marker to study the effect of labeling concentrations( 0. 2,2,20,50,100,200 μg / mL) and time( 1,15,30, 60,120,240 min) on the fluorescence intensity of V. anguillarum labeled FITC. In the meanw hile,the stability of fluorescence labeled on V. anguillarum stored at different temperatures( 25,4,- 20,- 70 ℃) and different time( 24,48,96,192 h) w ere compared. The possible change of fluorescence intensity w as monitored w hen V. anguillarum labeled FITC w as inactivated by formaldehyde. Also the fluorescence quenching effects of different intensity natural light( 0,150,1 500,25 000 lx) indoors and the effects of repeated freezing and thaw ing at- 20 and- 70 ℃ on V. anguillarum labeled FITC w ere analyzed. The results show ed that the optimal concentration of FITC w as in the range of 20 to 100 μg / mL and the optimal time w as in the range of 60 to 120 min. The fluorescent signal from V. anguillarum labeled FITC w as more stable w hen it w as stored at- 20 and- 70 ℃ than those stored at 4 and 25 ℃. The stability of fluorescent signal from formaldehyde inactivated V. anguillarum labeled FITC w as better than that from the control,and no difference w as found in the quenching effect in different intensity of natural lighting. After 3 times of freezing and thaw ing,the fluorescent signal from V. anguillarum labeled FITC frozen at- 70 ℃ w as more stable. The results above provided useful technical parameters w hen FITC w as used as a marker to label the marine bacteria.

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