光学学报, 2004, 24 (4): 536, 网络出版: 2006-06-12  

隐失驻波全内反射荧光显微术的数值计算

A Numerical Calculation of Super-Resolution Fluorescence Microscopy by Evanescent Standing Wave Illumination
作者单位
中国科学院上海光学精密机械研究所强光光学开放实验室, 上海 201800
摘要
从理论上详细计算了在全内反射的条件下,两束入射光产生的隐失干涉场(即隐失驻波)的强度分布,并分析了其不同于传统传播波干涉场的特点。同时使用数值模拟证明了利用隐失干涉场,即隐失驻波的激发方式,可以提高系统的分辨力,在横向实现超经典分辨的荧光成像。具体的分析表明,两束光以相等的角度入射,同时振幅相等,偏振态相同,所形成干涉条纹的反衬度最高,此时成像系统的有效点扩展函数最优化;入射介质1的折射率越大,隐失干涉场的空间周期越短(空间频率越高),其对应的调制点扩展函数中心瓣的半峰全宽越小,可能分辨更小的物体,但同时旁瓣的强度也增强,最终成像的分辨力受两者的共同制约。
Abstract
Standing wave total internal reflection imaging technique can obtain high resolution images of a sample along both axial and lateral directions. The variations of the interference patterns with incident condition of excitation light as well as refractive index of medium, and the differences between standing evanescent wave and standing propagating wave are theoretically investigated. The analysis shows that when contrast of the evanescent interference pattern is 1, optimal point spread function (PSF) can be obtained. Due to the narrowed FWHM of PSF, the lateral resolution increases with the decrease of period of the evanescent interference patterns. However, the intensity of PSF side bands increases, which will lower imaging quality.

王琛, 王桂英, 徐至展. 隐失驻波全内反射荧光显微术的数值计算[J]. 光学学报, 2004, 24(4): 536. 王琛, 王桂英, 徐至展. A Numerical Calculation of Super-Resolution Fluorescence Microscopy by Evanescent Standing Wave Illumination[J]. Acta Optica Sinica, 2004, 24(4): 536.

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