荧光共聚焦显微镜中一般用什么荧光探针

2025年03月15日 03:02
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荧光共聚焦显微镜中一般用什么荧光探针
目前常用的荧光探针有荧光素类探针、无机离子荧光探针、荧光量子点、分子信标等。荧光探针除应用于核酸和蛋白质的定量分析外,在核酸染色、DNA电泳、核酸分子杂交、定量PCR技术以及DNA测序上都有着广泛的应用。
目前,检测荧光探针的方法主要有单点测定和电荷耦合装置(CCD)荧光成像(包括用于微区分析的激光共聚焦荧光显微镜成像)。由于光电倍增管点扫描时间较长,激光照射强度高,很难抓住荧光早期变化。而CCD荧光成像的面阵大,成像视野广,成像时间可以调节,因而检测效果比较好。
化学发光检测的最大特点是设备简单、操作简便、分析速度快及灵敏度高。化学发光成像分析(CLI)是将化学发光与成像技术相结合,从而具有分辨率高、多样品同时检测、光谱响应范围宽以及灵敏度高等特点[10,11],已广泛应用于凝胶、蛋白印记及微阵列芯片中的化学发光信号检测。本实验建立了TCPO?咪唑?H2O2?荧光探针化学发光成像体系。由于化学发光不需要任何光源,因而在对荧光探针进行化学发光成像检测时,不存在荧光检测或者荧光成像时不可避免的光学背景的干扰,从而可以获得更低的检出限。用此体系对5种荧光探针进行定量分析,并研究了用四甲基异硫氰酸罗丹明(TRITC)标记的单克隆羊抗人IgG的化学发光成像,检出限达10-11mol/L,比相同条件下荧光成像的检出限低一个数量级。

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