Department of Molecular Biology and Center for Computational and Integrative Biology, Massachusetts General Hospital, Boston, Massachusetts.
Atomic and Molecular Physics Division, Harvard Smithsonian Center for Astrophysics, Cambridge, Massachusetts.
Biophys J. 2019 Feb 19;116(4):659-669. doi: 10.1016/j.bpj.2019.01.006. Epub 2019 Jan 10.
Having a fast, reliable method for characterizing vesicles is vital for their use as model cell membranes in biophysics, synthetic biology, and origins of life studies. Instead of the traditionally used Rayleigh-Gans-Debye approximation, we use an exact extended Lorenz-Mie solution for how core-shell particles scatter light to model vesicle turbidity. This approach enables accurate interpretations of simple turbidimetric measurements and is able to accurately model highly scattering vesicles, such as larger vesicles, those with multiple layers, and those with encapsulated material. We uncover several surprising features, including that vesicle lamellarity has a larger effect on sample turbidity than vesicle size and that the technique can be used to measure the membrane thickness of vesicles. We also examine potential misinterpretations of turbidimetry and discuss when measurements are limited by forward and multiple scattering and by the geometry of the instrument.
拥有一种快速、可靠的方法来描述囊泡对于将其作为生物物理、合成生物学和生命起源研究中的模型细胞膜非常重要。我们使用了精确的扩展 Lorenz-Mie 解来替代传统的瑞利-甘斯-德拜近似,以模拟囊泡的浊度。这种方法可以准确解释简单的比浊测量,并能够准确模拟高度散射的囊泡,如较大的囊泡、具有多层的囊泡和封装物质的囊泡。我们发现了一些令人惊讶的特征,包括囊泡的层状结构对样品浊度的影响比囊泡的大小更大,以及该技术可用于测量囊泡的膜厚度。我们还研究了比浊法可能存在的误解,并讨论了在哪些情况下测量受到前向和多次散射以及仪器几何形状的限制。