Comparative Analysis of Stain-Free Fourier Ptychographic Microscopy and Traditional Histopathological Light Microscopy in Renal Membranous Nephropathy

Marika Valentino, Vittorio Bianco, Gioacchino D'Ambrosio, Marco Paulli, Giovanni Smaldone, Valentina Brancato, Lisa Miccio, Marco Salvatore, Marcello Gambacorta, Pietro Ferraro
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Abstract

Background: Histology remains a cornerstone in the diagnosis and prognosis of renal diseases, with histopathological analysis of kidney tissue being crucial for understanding renal pathophysiology. The availability of multiple stained sections is essential for conducting a comprehensive histopathological analysis and achieving an accurate diagnosis. Recently, Fourier Ptychographic Microscopy (FPM) earned a spot among the most promising microscopy techniques. The ability to provide high-resolution, quantitative phase-contrast images over a wide area, particularly in a stain-free mode, makes FPM highly appealing to experts in histopathology. Since renal pathologies are characterized by subtle morphological changes encoded in tissue slides, phase maps obtained using FPM are well-suited for providing detailed, high-contrast images of tissue structures. Thus, FPM provides a quantitative imaging tool that can be descriptive of the sample and/or expressive of the disease. Methods: In this study, we explore FPM capability to image pathological kidney tissue, enabling pathologists to select regions of interest within the intricate architecture of renal tissue and zoom in to observe minute submicron structures, ranging from overall tissue organization and glomeruli distribution to individual cell membranes. Attention is focused on membranous glomerulonephritis since it is a nephropathy highly dependent on histological examination. Results: The comparative analysis between FPM and traditional light microscopy showed a difference in thickness of glomerular basal membranes between healthy kidney tissues and those affected by membranous glomerulonephritis (MG). Moreover, the results reported in our investigation revealed better glomerular membranes contrast in FPM images with respect to the H&E-stained images. Conclusions: Our study shows the broad potential of FPM in characterizing hallmarks of MG disease even in stain-free tissue slides.
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无染色傅立叶平片显微镜与传统组织病理学光镜在肾膜性肾病中的应用对比分析
背景:组织学仍然是肾脏疾病诊断和预后的基石,肾组织的组织病理学分析对于了解肾脏病理生理学至关重要。要进行全面的组织病理学分析并获得准确的诊断,必须有多个染色切片。最近,傅立叶层析显微镜(FPM)成为最有前途的显微镜技术之一。傅立叶相差显微镜能够提供大面积的高分辨率、定量相位对比图像,尤其是在无染色模式下,因此对组织病理学专家极具吸引力。由于肾脏病变的特征是组织切片中细微的形态变化,因此使用 FPM 获得的相位图非常适合提供组织结构的详细高对比度图像。因此,FPM 提供了一种定量成像工具,可以描述样本和/或表达疾病:在本研究中,我们探索了 FPM 对病理肾组织成像的能力,使病理学家能够在错综复杂的肾组织结构中选择感兴趣的区域,并放大观察微小的亚微米结构,从整体组织结构和肾小球分布到单个细胞膜。由于膜性肾小球肾炎是一种高度依赖组织学检查的肾病,因此重点关注膜性肾小球肾炎:结果:FPM 与传统光学显微镜的对比分析表明,健康肾组织与受膜性肾小球肾炎(MG)影响的肾组织之间的肾小球基底膜厚度存在差异。此外,我们的研究结果显示,与 H&E 染色图像相比,FPM 图像的肾小球膜对比度更高:我们的研究表明,即使在无染色的组织切片中,FPM 在描述 MG 疾病特征方面也具有广泛的潜力。
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