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Exploring the cellular and tissue uptake of nanomaterials in a range of biological samples using multimodal nonlinear optical microscopy

机译：使用多峰非线性光学显微镜探索一系列生物样品中纳米材料的细胞和组织吸收

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The uptake of nanomaterials (NMs) by cells is critical in determining their potential biological impact, whether beneficial or detrimental. Thus, investigation of NM internalization by cells is a common consideration in hazard and efficacy studies. There are currently a number of approaches that are routinely used to investigate NM-cell interactions, each of which have their own advantages and limitations. Ideally, imaging modalities used to investigate NM uptake by cells should not require the NM to be labelled (e.g. with fluorophores) to facilitate its detection. We present a multimodal imaging approach employing a combination of label-free microscopies that can be used to investigate NM-cell interactions. Coherent anti-Stokes Raman scattering microscopy was used in combination with either two-photon photoluminescence or four-wave mixing (FWM) to visualize the uptake of gold or titanium dioxide NMs respectively. Live and fixed cell imaging revealed that NMs were internalized by J774 macrophage and C3A hepatocyte cell lines (15-31 mu g ml(-1)). Sprague Dawley rats were exposed to NMs (intratracheal instillation, 62 mu g) and NMs were detected in blood and lung leucocytes, lung and liver tissue, demonstrating that NMs could translocate from the exposure site. Obtained data illustrate that multimodal nonlinear optical microscopy may help overcome current challenges in the assessment of NM cellular uptake and biodistribution. It is therefore a powerful tool that can be used to investigate unlabelled NM cellular and tissue uptake in three dimensions, requires minimal sample preparation, and is applicable to live and fixed cells.
机译：细胞对纳米材料（NMs）的吸收对于确定其潜在的生物学影响（无论有益还是有害）至关重要。因此，在危害和功效研究中，细胞对NM内在化的研究是普遍考虑的问题。当前，有许多常规用于研究NM细胞相互作用的方法，每种方法都有其自身的优点和局限性。理想地，用于研究细胞对NM吸收的成像方式不应要求标记NM（例如用荧光团）以促进其检测。我们提出了一种多模态成像方法，该方法采用了无标记显微技术的组合，可用于研究NM细胞相互作用。相干抗斯托克斯拉曼散射显微镜与两光子光致发光或四波混合（FWM）结合使用，分别观察了金或二氧化钛NM的吸收。实时和固定细胞成像显示，NMs被J774巨噬细胞和C3A肝细胞系（15-31μg ml（-1））内化。将Sprague Dawley大鼠暴露于NMs（气管内滴注，62微克），并且在血液和肺白细胞，肺和肝组织中检测到NMs，表明NMs可以从暴露部位转移。获得的数据表明，多峰非线性光学显微镜可能有助于克服目前对NM细胞摄取和生物分布评估的挑战。因此，它是一个功能强大的工具，可用于在三个维度上研究未标记的NM细胞和组织的摄取，需要最少的样品制备，并且适用于活细胞和固定细胞。

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