Atomic force microscopy (AFM) is a consolidated technique for the study of biological systems, usually ex vivo or in culture, under different experimental conditions. Yet, the diffusion of the technique in the scientific context of histology is still rather slow and limited. In the present work, we demonstrate the potential of AFM, in terms of morphological and nanomechanical imaging, to study the effects of nano‐ and micro‐sized metallic pollutants in living biological systems. As a model, we investigated marine molluscs (Mytilus galloprovincialis) grown in the Adriatic Sea. We characterized histological sections from two organs (gonads and digestive glands) of molluscs collected during several surveys at different growth time and distance from gas extraction platforms. We evaluated the effects of nano‐pollutants mostly on the local tissue structure by combining AFM microscopy with scanning electron microscopy (SEM). Furthermore, the AFM images allowed evidencing the presence of nano‐ or micro‐sized structures that exhibit different nanomechanical properties compared to the rest of the tissue. The results demonstrate how coupling AFM and SEM analysis can provide an effective procedure to evaluate the morphological alterations produced by the exposure to exogenous nano‐pollutants in tissue and constitute a promising way to reveal basic mechanisms mediating the cytotoxicity of specific exogenous pollutants ingested by edible organisms.

中文翻译：

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海洋生物中金属基微纳米污染物：我们可以从原子力显微镜-扫描电子显微镜联合研究中学到什么

原子力显微镜 (AFM) 是一种综合技术，用于研究不同实验条件下的生物系统，通常是离体或培养的生物系统。然而，该技术在组织学科学背景下的传播仍然相当缓慢和有限。在目前的工作中，我们展示了 AFM 在形态学和纳米力学成像方面的潜力，用于研究纳米和微米级金属污染物对生物系统的影响。作为模型，我们研究了海洋软体动物（Mytilus galloprovincialis) 生长在亚得里亚海。我们对在不同生长时间和距气体提取平台距离的几次调查中收集的软体动物的两个器官（性腺和消化腺）的组织切片进行了表征。我们通过结合 AFM 显微镜和扫描电子显微镜 (SEM) 来评估纳米污染物主要对局部组织结构的影响。此外，AFM 图像可以证明纳米级或微米级结构的存在，与组织的其余部分相比，这些结构表现出不同的纳米力学特性。