Comprehensive analysis of the multifractional molecular diffusion provides a deeper understanding of the diffusion phenomenon in the fields of material science, molecular and cell biology, advanced biomaterials, etc. Fluorescence recovery after photobleaching (FRAP) is commonly employed to probe the molecular diffusion. Despite FRAP being a very popular method, it is not easy to assess multifractional molecular diffusion due to limited possibilities of approaches for analysis. Here we present a novel simulation-optimization-based approach (S-approach) that significantly broadens possibilities of the analysis. In the S-approach, possible fluorescence recovery scenarios are primarily simulated and afterward compared with a real measurement while optimizing parameters of a model until a sufficient match is achieved. This makes it possible to reveal multifractional molecular diffusion. Fluorescent latex particles of different size and fluorescein isothiocyanate in an aqueous medium were utilized as test systems. Finally, the S-approach has been used to evaluate diffusion of cytochrome c loaded into multilayers made of hyaluronan and polylysine. Software for evaluation of multifractional molecular diffusion by S-approach has been developed aiming to offer maximal versatility and user-friendly way for analysis.

中文翻译：

---

基于FRAP的聚合物多层分子扩散的多部分分析：一种基于模拟的新方法

对多部分分子扩散的全面分析提供了对材料科学，分子和细胞生物学，先进生物材料等领域中扩散现象的更深入了解。光漂白后的荧光恢复（FRAP）通常用于探测分子扩散。尽管FRAP是一种非常流行的方法，但是由于分析方法的可能性有限，因此评估多组分分子扩散并不容易。在这里，我们提出了一种新颖的基于仿真优化的方法（S方法），该方法大大拓宽了分析的可能性。在S方法中，首先对可能的荧光回收方案进行了模拟，然后与实际测量值进行了比较，同时优化了模型的参数，直到获得足够的匹配为止。这使得揭示多组分分子扩散成为可能。将不同尺寸的荧光胶乳颗粒和水性介质中的异硫氰酸荧光素用作测试系统。最后，S方法已用于评估加载到透明质酸和聚赖氨酸制成的多层膜中的细胞色素c的扩散。开发了用于通过S方法评估多分子扩散的软件，旨在提供最大的多功能性和用户友好的分析方法。