激光与光电子学进展, 2014, 51 (1): 011102, 网络出版: 2013-12-20  

偏振态和有效数值孔径对共聚焦全内反射显微术分辨率的影响 下载: 624次

Effects of Polarization State and Effective Numerical Aperture on the Resolution in Confocal Total Internal Reflection Microscopy
魏通达 1,2,3,*张运海 3肖昀 1,2,3唐玉国 1,3
作者单位
1 中国科学院长春光学精密机械与物理研究所, 吉林 长春 130033
2 中国科学院大学, 北京 100049
3 中国科学院苏州生物医学工程技术研究所, 江苏 苏州 215163
摘要
为了研究入射光偏振态和入射光瞳有效数值孔径对共聚焦全内反射显微术(CTIRM)的分辨率,即焦点横向半峰全宽和纵向倏逝场透射深度的影响,根据Richard-Wolf矢量衍射理论,计算并讨论了线偏振、圆偏振、径向偏振和切向偏振入射光在界面处的光强分布情况;同时计算了在三种不同的有效数值孔径(1.33~1.45,1~1.45和1~1.12)时,焦点半峰全宽和倏逝场透射深度的数值结果。计算结果表明,当有效数值孔径为1.33~1.45,入射波长为532 nm时,径向偏振光在横向上可达144 nm的半峰全宽,优于线偏振的330 nm和圆偏振的360 nm,超过了衍射极限。薄且大的有效数值孔径能够获得更小的半峰全宽,其上下限的平方差越大,透射深度越小。三种数值孔径中,1~1.45时的透射深度最小,为140 nm。相比较其他偏振光,径向偏振光更适合作为共聚焦全内反射显微术的入射激发光,并能够通过优化有效数值孔径,获得样品近表面处的高横向分辨率和低轴向荧光噪声。
Abstract
In order to study the effects of polarization state and effective numerical aperture on the focal full-width at half-maximum (FWHM) and evanescent depth in confocal total internal reflection Microscopy(CTIRM), according to the Richard-Wolf theory, the intensity distribution in the interface is calculated and discussed with linearly, circularly, radially and azimuthally polarized beams, respectively. Meanwhile, the numerical calculations of FWHM and depth are researched for three different effective numerical apertures (1.33~1.45, 1~1.45 and 1~1.12, respectively). The results indicate that the FWHM of radially polarized beam is 144 nm, breaking through the limitation of diffraction, which is also better than 330 nm of linearly and 360 nm of circularly polarized ones, by using 532 nm incident wave and 1.33~1.45 aperture. The sharper focuscan be got with the larger and thinner aperture. Also the smaller depth of transmission can be got with the larger difference of squares of upper and lower limits, for which 140 nm is the shallowest depth with 1~1.45 aperture. Compared to other polarizations, radially polarized beam is the most suitable choice for CTIRM. And by optimizing the effective numerical aperture, high horizontal resolutions and low axial fluorescent noise near the sample surfacecan be obtained simultaneously.

魏通达, 张运海, 肖昀, 唐玉国. 偏振态和有效数值孔径对共聚焦全内反射显微术分辨率的影响[J]. 激光与光电子学进展, 2014, 51(1): 011102. Wei Tongda, Zhang Yunhai, Xiao Yun, Tang Yuguo. Effects of Polarization State and Effective Numerical Aperture on the Resolution in Confocal Total Internal Reflection Microscopy[J]. Laser & Optoelectronics Progress, 2014, 51(1): 011102.

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