Removing the effect of blooming from potential energy measurement by employing total internal reflection microscopy integrated with video microscopy
Removing the effect of blooming from potential energy measurement by employing total internal reflection microscopy integrated with video microscopy
复制标题
通过采用与视频显微镜集成的全内反射显微镜消除势能测量中的光晕影响
DOI：
10.1016/j.jcis.2017.05.024
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发表时间：
2017
影响因子：
9.9
通讯作者：
Ngai To
中科院分区：
文献类型：
--
作者：
Cao Feng;Gong Xiangjun;He Chuanxin;Ngai To
全内反射显微镜(TIRM)测量了水溶液中胶体颗粒与平面之间的相互作用。最近，TIRM进一步与视频显微镜(VM)相结合，能够同时测量同一系综中的多粒子胶体-表面相互作用。然而，由于图像的蓬勃发展，精确的图像获取仍然存在挑战。光晕意味着到达探测器的光子数量超过了它的最大容量，多余的光子要么溢出到相邻的像素，要么不被计算，导致无法准确确定强度。我们的结果表明，光晕引起了测量的势能与经典的Derjaguin，Landau，Verway和Overbeek(DLVO)理论的偏差。因此，本文提出了一种从实验测量中推断出真实光强的修正方法。在光晕发生时，散射光强度与曝光时间的关系偏离线性关系。建立了恢复真实光强的修正方程，并通过相应的势能分布与DLVO理论的一致性进行了验证。这种校正方法适用于散射光照射的胶体探针的VM系统，拓宽了VM成像在胶体相互作用研究中的应用。
Total internal reflection microscopy (TIRM) measures the interactions between a colloidal particle and a flat surface in aqueous solution. Recently, TIRM has further integrated with video microscopy (VM) and enabled the simultaneous measurements of multi-particle colloid-surface interactions in the same ensemble. However, there still remain challenges about accurate image acquisition due to blooming. Blooming means the number of photons reaching the detector exceeds its maximum capacity, and the excess photons will either spill to adjacent pixels or not be counted, leading to an obstacle from precise determination of intensity. Our result shows that blooming gives rise to a deviation of the measured potential energy from the classical theory of Derjaguin, Landau, Verway, and Overbeek (DLVO). Therefore, a correction method was developed in this work to deduce the real intensity from the experimental measurement. The relationship between scattered light intensity and exposure time deviates from linearity when blooming occurs. A correction equation was developed to recover the real intensity, which was then confirmed by the accordance between the corresponding potential energy profiles and the DLVO theory. This correction method is suitable for VM systems of colloidal probes illuminated by scattered light, broadening the application of VM imaging to investigate colloidal interactions.