论文标题

使用荧光珠模拟单flurophores

Using fluorescent beads to emulate single flurophores

论文作者

Aleman-Castaneda, Luis A., Feng, Sherry Yi-Ting, Gutierrez-Cuevas, Rodrigo, Herrera, Isael, Brown, Thomas G., Brasselet, Sophie, Alonso, Miguel A.

论文摘要

在这项工作中,我们研究了在光学显微镜校准中可以使用荧光珠模拟单个荧光分子的条件。尽管珠子由于其亮度和易于操作而被广泛使用,但它们产生的点扩散函数(PSF)与单分子荧光团的分量函数之间可能存在明显的差异,这是由于其不同的发射模式和大小而引起的。我们从理论上研究各种情况的这些差异,例如有或没有极化通道分裂,以确定将珠子用作单分子模型的条件是有效的。我们还提出了由于尺寸差异而模拟模糊的方法,并补偿了它,以产生与单分子更相似的PSF。

In this work, we study the conditions under which fluorescent beads can be used to emulate single fluorescent molecules in the calibration of optical microscopes. Although beads are widely used due to their brightness and easy manipulation, there can be notable differences between the point spread functions (PSFs) they produce and those for single-molecule fluorophores, caused by their different emission pattern and their size. We study theoretically these differences for various scenarios, e.g. with or without polarization channel splitting, to determine the conditions under which the use of beads as a model for single molecules is valid. We also propose methods to model the blurring due to the size difference and compensate for it to produce PSFs that are more similar to those for single molecules.

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