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自体荧光是生物体内部组织成分在吸收光时自然发出的荧光,会影响人工标记荧光的有效识别,严重干扰对抗体标记的目的蛋白荧光的观察及数据分析,在珊瑚冰冻切片免疫荧光标记研究中尤为突显。为更有效地去除珊瑚组织切片中的自体荧光,本研究以共生模式种花伞软珊瑚（Xenia sp.）为研究对象,首次尝试在花伞软珊瑚冰冻切片免疫荧光染色中,使用一种在小鼠和人体组织切片中常用的TrueBlack Lipofuscin自体荧光淬灭剂,并与已有研究中的酒精梯度脱水法进行比较。结果表明,使用荧光淬灭剂TrueBlack Lipofuscin后,花伞软珊瑚组织切片中的自体荧光强度是传统的酒精梯度脱水法的52.4%,且在Cy5通道下效果最佳。同时,通过珊瑚免疫荧光标记实验表明使用TrueBlack Lipofuscin的实验组特异性荧光信号强度为对照组的205.3%,背景荧光和自体荧光基本去除。 相似文献
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SHENG Xiuzhen ZHAN Wenbin XU Songjuan CHENG Shunfeng 《中国海洋大学学报(英文版)》2007,6(4):378-382
Lymphocystis nodules occurring in the cultured sting fish Sebastes schlegeli were observed under light and electron microscope. Lymphocystis disease virus (LCDV) in the tissues of diseased fish was detected with indirect immunofluorescence test (IFAT). Results showed that lymphocystis cells had overly irregular nuclei, basophilic intracytoplasmic inclusion bodies with virions budding from the surface, and hyaline capsules outside the cell membrane. Numerous virus particles about 200 nm in diameter scat- tered in the cytoplasm, electron-dense particles 70-80 nm in diameter filled in perinuclear cisterna, and membrane-enveloped parti- cles with electron-dense core of 70-80 nm appeared around cellular nucleus. IFAT using monoclonal antibody against LCDV from Paralichthys olivaceus revealed that specific green fluorescence was present in the cytoplasm of lymphocystis cells, epithelium of stomach, gill lamellae, and muscular fibers under epidermis of S. schlegeli, just as that in the cytoplasm of lymphocystis cells of P. olivaceus, suggesting the presence of LCDV in these tissues. 相似文献
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白斑综合症病毒囊膜蛋白VP28单链抗体的酵母表面展示及其抗病毒特性的初步研究 总被引：1，自引：0，他引：1
白斑综合症病毒(white spot syndrome virus,WSSV)是对虾养殖业中危害最大的病原微生物之一,对对虾养殖业造成了巨大的经济损失,但目前尚无有效防治手段.本研究从WSSV囊膜蛋白VP28免疫的小鼠细胞中分别扩增到重链可变区基因和轻链可变区基因,大小为351 bp和342 bp.两基因序列通过Linker序列连接后,构建到酵母表面载体pYD1中,得到重组质粒pYD1-vp28scfv.将该重组质粒在酿酒酵母EBY100中诱导表达,通过免疫荧光实验发现,重组蛋白VP28scFv在酿酒酵母EBY100表面实现展示.进一步通过结合实验表明该展示蛋白具有WSSV的结合活性.本研究在结合酿酒酵母可作为饲料添加剂的特性基础上,对WSSV囊膜蛋白VP28单链抗体的抗病毒特性进行了应用,从而为WSSV的防治提供了新思路. 相似文献
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