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基于径向偏振光束的全内反射荧光显微成像具有更高的成像分辨率,探究其倏逝场的强度分布规律是实现系统结构设计和优化的关键。基于矢量衍射理论,推导了聚焦的径向偏振倏逝场的数学表达式,进行了理论计算,并基于扫描近场光学显微镜搭建了实验系统,测试了倏逝场的强度分布,验证了理论分析。测试结果表明,聚焦的径向偏振光束产生的倏逝场在横向及轴向都具有非常高的分辨率,在可见光波长范围内,光斑横向半高全宽度在100~200nm范围,轴向半高全宽度在40~80nm范围,突破了衍射极限,证明了该类型倏逝场在全内反射荧光显微成像领域巨大的应用价值,也为接下来的系统改进提供了技术支持。
Total internal reflection fluorescence microscopy imaging based on radial polarized beam has higher imaging resolution, and exploring the law of intensity distribution of evanescent field is the key to realize system structure design and optimization. Based on the vector diffraction theory, the mathematical expression of the focused radial polarization evanescent field was deduced, and the theoretical calculation was carried out. The experimental system was built based on the scanning near-field optical microscope and the intensity distribution of the evanescent field was tested. The theoretical analysis . The test results show that the evanescent field generated by the focused radial polarized beam has a very high resolution both in the transverse direction and the axial direction. In the visible wavelength range, the full width at half maximum of the transverse spot width is in the range of 100-200 nm and the axial half height The full width in the range of 40 ~ 80nm, breaking the diffraction limit, proved that this type of evanescent field in the field of total internal reflection fluorescence microscopy of great value, but also for the subsequent system improvements provide technical support.