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首页> 美国卫生研究院文献>Journal of Biomedical Optics >Angularly resolved finely sampled elastic scattering measurements of single cells: requirements for robust organelle size extractions
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Angularly resolved finely sampled elastic scattering measurements of single cells: requirements for robust organelle size extractions

机译：角分辨精细采样的单细胞弹性散射测量：稳健的细胞器大小提取的要求

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摘要

Angularly resolved elastic light scattering is an established technique for probing the average size of organelles in biological tissue and cellular ensembles. Focusing of the incident light to illuminate no more than one cell at a time restricts the minimum forward-scattering angle that can be detected. Series of simulated single-cell angular-scattering patterns have been generated to explore how size estimates vary as a function of . At a setting of , the size estimates hop unstably between multiple minima in the solution space as simulated noise (mimicking experimentally observed levels) is varied. As is reduced from 20 deg to 10 deg, the instability vanishes, and the variance of estimates near the correct answer also decreases. The simulations thus suggest that robust Mie theory fits to single-cell scattering at 785 nm excitation require measurements down to at least 15 deg. Notably, no such instability was observed at for narrow bead distributions. Accurate sizing of traditional calibration beads is, therefore, insufficient proof that an angular-scattering system is capable of robust analysis of single cells. Experimental support for the simulation results is also presented using measurements on cells fixed with formaldehyde.
机译：角分辨弹性光散射是一种探测生物组织和细胞团中细胞器平均大小的成熟技术。入射光的聚焦一次最多照亮一个单元，限制了可以检测到的最小前向散射角。已生成一系列模拟的单细胞角散射图案，以探索尺寸估计值如何随的变化。在的设置下，随着模拟噪声（模仿实验观察到的水平）的变化，大小估计在解决方案空间中的多个最小值之间不稳定地跳跃。当从20度减小到10度时，不稳定性消失，并且正确答案附近的估计方差也减小了。因此，仿真表明健壮的Mie理论适合于785 nm激发下的单细胞散射，要求至少进行15度的测量。值得注意的是，对于窄的珠粒分布，没有观察到这种不稳定性。因此，传统校准珠的准确尺寸确定不足以证明角度散射系统能够对单个细胞进行可靠的分析。还通过使用甲醛固定的电池进行测量来提供对模拟结果的实验​​支持。

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