A quantitative description for optical mass measurement of single biomolecules
A quantitative description for optical mass measurement of single biomolecules
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单个生物分子光学质量测量的定量描述
DOI：
10.1101/2023.03.28.534430
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Becker J
中科院分区：
文献类型：
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作者：
Becker J
在过去的十年里，使用各种实验方法已经实现了对溶液中单个生物分子的无标记检测。然而，我们对光学对比度的大小及其与基本原子结构的关系以及可实现的测量灵敏度和精度的理解仍然不够明确。在这里，我们使用傅里叶光学方法结合基于原子结构的分子极化率模型，从第一性原理模拟质量光度学实验。我们发现几个关键的实验确定的参数，如光学对比度到质量的转换，可实现的质量精度，以及与分子形状和取向的关系，都有很好的一致性。这使我们能够确定检测灵敏度和测量精度，在很大程度上独立于所选择的光学检测方法，从而产生基于光的单分子检测和定量的一般框架。
Label-free detection of single biomolecules in solution has been achieved using a variety of experimental approaches over the past decade. Yet, our understanding of the magnitude of the optical contrast and its relationship with the underlying atomic structure as well as the achievable measurement sensitivity and precision remain poorly defined. Here, we use a Fourier optics approach combined with an atomic structure-based molecular polarizability model to simulate mass photometry experiments from first principles. We find excellent agreement between several key experimentally determined parameters such as optical contrast-to-mass conversion, achievable mass accuracy, and molecular shape and orientation dependence. This allows us to determine detection sensitivity and measurement precision mostly independent of the optical detection approach chosen, resulting in a general framework for light-based single-molecule detection and quantification.