Morgridge Institute for Research, Madison, Wisconsin, United States.
Colorado State University, Fort Collins, Colorado, United States.
J Biomed Opt. 2024 Jun;29(Suppl 2):S22716. doi: 10.1117/1.JBO.29.S2.S22716. Epub 2024 Sep 5.
The emergence of label-free microscopy techniques has significantly improved our ability to precisely characterize biochemical targets, enabling non-invasive visualization of cellular organelles and tissue organization. However, understanding each label-free method with respect to the specific benefits, drawbacks, and varied sensitivities under measurement conditions across different types of specimens remains a challenge.
We link all of these disparate label-free optical interactions together and compare the detection sensitivity within the framework of statistical estimation theory.
To achieve this goal, we introduce a comprehensive unified framework for evaluating the bounds for signal detection with label-free microscopy methods, including second-harmonic generation, third-harmonic generation, coherent anti-Stokes Raman scattering, coherent Stokes Raman scattering, stimulated Raman loss, stimulated Raman gain, stimulated emission, impulsive stimulated Raman scattering, transient absorption, and photothermal effect. A general model for signal generation induced by optical scattering is developed.
Based on this model, the information obtained is quantitatively analyzed using Fisher information, and the fundamental constraints on estimation precision are evaluated through the Cramér-Rao lower bound, offering guidance for optimal experimental design and interpretation.
We provide valuable insights for researchers seeking to leverage label-free techniques for non-invasive imaging applications for biomedical research and clinical practice.
无标记显微镜技术的出现极大地提高了我们精确描述生化靶标的能力，使细胞器官和组织结构的非侵入可视化成为可能。然而，了解每种无标记方法相对于不同类型标本在测量条件下的具体优点、缺点和不同灵敏度仍然是一个挑战。
我们将所有这些不同的无标记光相互作用联系在一起，并在统计估计理论的框架内比较检测灵敏度。
为了实现这一目标，我们引入了一个全面的统一框架，用于评估无标记显微镜方法的信号检测的界限，包括二次谐波产生、三次谐波产生、相干反斯托克斯拉曼散射、相干斯托克斯拉曼散射、受激拉曼损耗、受激拉曼增益、受激辐射、脉冲受激拉曼散射、瞬态吸收和光热效应。开发了一种用于光散射引起的信号产生的通用模型。
基于该模型，使用 Fisher 信息对获得的信息进行定量分析，并通过 Cramér-Rao 下限评估估计精度的基本约束，为最佳实验设计和解释提供指导。
我们为寻求利用无标记技术进行生物医学研究和临床实践中非侵入性成像应用的研究人员提供了有价值的见解。
