Spatio-angular fluorescence microscopy III. Constrained angular diffusion, polarized excitation, and high-NA imaging.
Spatio-angular fluorescence microscopy III. Constrained angular diffusion, polarized excitation, and high-NA imaging.
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DOI：
10.1364/josaa.389217
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发表时间：
2020-09-01
期刊：
影响因子：
--
通讯作者：
La Rivière P
中科院分区：
文献类型：
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作者：
Chandler T;Shroff H;Oldenbourg R;La Rivière P
我们研究了荧光分子在角势阱中的旋转扩散，这些扩散分子的激发和随后的发射，以及用高NA无光晕光学显微镜对这些发射的成像。虽然偶极辐射只传输六个低频角分量，但我们发现角结构照明可以将高频角分量混叠到成像系统的通带中。结果表明，旋转扩散时间、荧光衰减时间和采集时间这三个时间尺度之间的关系限制了可测角度分量的数量。我们通过在快角扩散、慢角扩散和弱势的限制下模拟一个数值体模来演示我们的模型。
We investigate rotational diffusion of fluorescent molecules in angular potential wells, the excitation and subsequent emissions from these diffusing molecules, and the imaging of these emissions with high-NA aplanatic optical microscopes. Although dipole emissions only transmit six low-frequency angular components, we show that angular structured illumination can alias higher-frequency angular components into the passband of the imaging system. We show that the number of measurable angular components is limited by the relationships between three time scales: the rotational diffusion time, the fluorescence decay time, and the acquisition time. We demonstrate our model by simulating a numerical phantom in the limits of fast angular diffusion, slow angular diffusion, and weak potentials.