Imaging of fluorescent polymer dots in relation to channels and immune cells in the lymphatic system.
Imaging of fluorescent polymer dots in relation to channels and immune cells in the lymphatic system.
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DOI：
10.1016/j.mtbio.2022.100317
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发表时间：
2022-06
影响因子：
8.2
通讯作者：
Xiong, Liqin
中科院分区：
文献类型：
--
作者：
Zhang, Yufan;Zhang, Juxiang;Li, Xiaowei;Li, Jingru;Lu, Shuting;Li, Yuqiao;Ren, Panting;Zhang, Chunfu;Xiong, Liqin
聚合物点（Pdot）已被应用于活体小鼠和大鼠的淋巴结（LN）和淋巴管（LV）成像。然而，Pdot在LN和LV中的吸收、分布、代谢和排泄机制仍不清楚。因此，本研究采用多种体内、体外显微成像和检测技术，研究了Pdot与免疫细胞、淋巴细胞和胶原纤维的关系。流式细胞术显示Pdots可被巨噬细胞和单核细胞吞噬，与淋巴结内的B细胞、T细胞和树突状细胞无关。银染、免疫荧光和双光子显微镜观察显示Pdot聚集于LN的胶原纤维和LV内。此外，免疫荧光成像结果证实Pdot分布在收集的LV内皮细胞的细胞外间隙。此外，在注射21天后，收集的LV中的Pdot基本上通过泄漏到周围组织或引流LN而被清除。在长期观察中，Pdots还有助于监测左室收缩频率和变化范围。本研究为以后以Pdots为载体研究淋巴管的结构和功能奠定了基础。我们设计并合成了五种Pdot，并通过多种体内、体外显微成像和检测技术研究了Pdot与免疫细胞或免疫通道的关系。在淋巴管（LV）中，Pdot主要分布于淋巴管内皮细胞的细胞外，并持续至21 d。此外，Pdots在LV、网状网络和吞噬细胞的帮助下对整个淋巴结（LN）进行成像。我们的研究为Pdot提供了一个成像时间窗口，以监测LN和LV的变化。
Polymer dots (Pdots) have been applied to imaging lymph nodes (LNs) and lymphatic vessels (LVs) in living mice and rats. However, the mechanism of absorption, distribution, metabolism, and excretion of Pdots in LNs and LVs is still unclear. Therefore, the relationship between Pdots and immune cells, LVs and collagen fibers in lymphatics was studied by multiple in vivo and ex vivo microscopic imaging methods and detection techniques. Flow cytometry showed that Pdots could be phagocytosed by macrophages and monocytes, and had no relationship with B cells, T cells and dendric cells in LNs. Silver staining, immunofluorescence and two-photon microscope showed that Pdots gathered in collagen fibers and LVs of LNs. Furthermore, immunofluorescence imaging results verified that Pdots were distributed in the extracellular space of collecting LVs endothelial cells. In addition, Pdots in the collecting LVs were basically cleared by leaking into the surrounding tissue or draining LNs after 21 days of injection. During the long-time observation, Pdots also helped monitor the contraction frequency and variation range of LV. Our study lays a foundation on the research of Pdots as the carrier to study lymphatic structure and function in the future. We designed and synthesized five Pdots, and studied the relationship between Pdots and immune cells or channels in lymphatics by multiple in vivo and ex vivo microscopic imaging methods and detection techniques. In the lymphatic vessels (LV), Pdots existed in the extracellular of lymphatic endothelial cells, and last up to 21 d. In addition, Pdots image the whole lymph node (LN) with the aid of LVs, reticular network, and phagocytosis cells. Our study provides an imaging-time window for Pdots to monitor the change both in the LNs and LVs.
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