A Quantitative Description for Optical Mass Measurement of Single Biomolecules.
A Quantitative Description for Optical Mass Measurement of Single Biomolecules.
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DOI：
10.1021/acsphotonics.3c00422
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发表时间：
2023-08-16
期刊：
影响因子：
7
通讯作者：
Kukura, Philipp
中科院分区：
文献类型：
--
作者：
Becker, Jan;Peters, Jack S.;Crooks, Ivor;Helmi, Seham;Synakewicz, Marie;Schuler, Benjamin;Kukura, Philipp
在过去的十年中，通过各种实验方法已经实现了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底层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明确。在这里，我们使用傅立叶光学方法与基于原子结构的分子极化率模型相结合，从第一原理模拟质量光度测定实验。我们发现几个关键的实验确定的参数之间非常一致，例如光学对比质量转换、可实现的质量精度以及分子形状和方向依赖性。这使我们能够确定检测灵敏度和测量精度，基本上与所选的光学检测方法无关，从而形成基于光的单分子检测和定量的通用框架。
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.
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