van Staalduine Simon E, Bianco Vittorio, Ferraro Pietro, Menzel Miriam
Department of Imaging Physics, Faculty of Applied Sciences, Delft University of Technology, Delft, the Netherlands.
Institute of Applied Sciences and Intelligent Systems, Italian National Research Council, Pozzuoli (Napoli), Italy.
bioRxiv. 2025 Apr 9:2024.11.28.625428. doi: 10.1101/2024.11.28.625428.
Fourier Ptychographic Microscopy (FPM) provides high-resolution imaging and morphological information over large fields of view, while Computational Scattered Light Imaging (ComSLI) excels at mapping interwoven fiber organization in unstained tissue sections. This study introduces Fourier Ptychographic Scattered Light Microscopy (FP-SLM), a new multi-modal approach that combines FPM and ComSLI analyses to create both high-resolution phase-contrast images and fiber orientation maps from a single dataset. The method is demonstrated on a state-of-the-art setup that was originally used for FPM and one that was originally used for ComSLI, and the outputs are quantitatively compared to each other on brain sections (frog, monkey) and sections from thigh muscle and knee (mouse). FP-SLM delivers high-resolution images while revealing fiber organization in nerve, muscle, tendon, cartilage, and bone tissues. The approach is validated by comparing the computed fiber orientations with those derived from structure tensor analysis of the high-resolution images. The comparison shows that FPM and ComSLI are compatible with each other and yield fully consistent results. Remarkably, this combination surpasses the sum of its parts, so that applying ComSLI analysis to FPM recordings and vice-versa outperforms both methods alone. FP-SLM can be retrospectively applied to analyze any existing dataset acquired from a setup that was originally built for FPM or ComSLI alone (consisting of LED array and low numerical aperture), without need to build or design an extra setup. This significantly expands the application range of both techniques and enhances the study of complex tissue architectures in biomedical research.
傅里叶叠层显微镜（FPM）可在大视野范围内提供高分辨率成像和形态学信息，而计算散射光成像（ComSLI）则擅长绘制未染色组织切片中交织的纤维组织。本研究介绍了傅里叶叠层散射光显微镜（FP-SLM），这是一种新的多模态方法，它结合了FPM和ComSLI分析，可从单个数据集中创建高分辨率相衬图像和纤维取向图。该方法在最初用于FPM的先进装置和最初用于ComSLI的装置上进行了演示，并在脑切片（青蛙、猴子）以及大腿肌肉和膝盖（小鼠）切片上对输出结果进行了定量比较。FP-SLM在揭示神经、肌肉、肌腱、软骨和骨组织中的纤维组织的同时，还能提供高分辨率图像。通过将计算得到的纤维取向与从高分辨率图像的结构张量分析中得出的纤维取向进行比较，验证了该方法的有效性。比较结果表明，FPM和ComSLI相互兼容，且产生的结果完全一致。值得注意的是，这种组合所达到的效果超过了其各部分单独作用的总和，因此将ComSLI分析应用于FPM记录，反之亦然，其性能优于单独使用这两种方法。FP-SLM可以追溯应用于分析从最初仅为FPM或ComSLI构建的装置（由LED阵列和低数值孔径组成）获取的任何现有数据集，而无需构建或设计额外的装置。这显著扩展了这两种技术的应用范围，并加强了生物医学研究中对复杂组织结构的研究。
