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首页> 美国卫生研究院文献>Journal of Visualized Experiments : JoVE >Real-time Imaging of Myeloid Cells Dynamics in ApcMin/+ Intestinal Tumors by Spinning Disk Confocal Microscopy
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Real-time Imaging of Myeloid Cells Dynamics in ApcMin/+ Intestinal Tumors by Spinning Disk Confocal Microscopy

机译：旋转盘共聚焦显微镜在ApcMin / +肠肿瘤中的髓样细胞动力学的实时成像。

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摘要

Myeloid cells are the most abundant immune cells within tumors and have been shown to promote tumor progression. Modern intravital imaging techniques enable the observation of live cellular behavior inside the organ but can be challenging in some types of cancer due to organ and tumor accessibility such as intestine. Direct observation of intestinal tumors has not been previously reported. A surgical procedure described here allows direct observation of myeloid cell dynamics within the intestinal tumors in live mice by using transgenic fluorescent reporter mice and injectable tracers or antibodies. For this purpose, a four-color, multi-region, micro-lensed spinning disk confocal microscope that allows long-term continuous imaging with rapid image acquisition has been used. ApcMin/+ mice that develop multiple adenomas in the small intestine are crossed with c-fms-EGFP mice to visualize myeloid cells and with ACTB-ECFP mice to visualize intestinal epithelial cells of the crypts. Procedures for labeling different tumor components, such as blood vessels and neutrophils, and the procedure for positioning the tumor for imaging through the serosal surface are also described. Time-lapse movies compiled from several hours of imaging allow the analysis of myeloid cell behavior in situ in the intestinal microenvironment.
机译：髓样细胞是肿瘤内最丰富的免疫细胞，并已显示出可促进肿瘤进展。现代的活体成像技术可以观察器官内部的活细胞行为，但由于器官和肿瘤的可及性（如肠道），在某些类型的癌症中可能具有挑战性。以前没有直接观察肠道肿瘤的报道。通过使用转基因荧光报告基因小鼠和可注射的示踪剂或抗体，本文所述的外科手术程序可以直接观察活体小鼠肠道肿瘤内的髓样细胞动力学。为此，已经使用了四色，多区域，微透镜旋转盘共聚焦显微镜，该显微镜可以进行连续的连续成像并具有快速的图像采集功能。在小肠中形成多个腺瘤的Apc Min / + 小鼠与c-fms-EGFP小鼠杂交以观察髓样细胞，并与ACTB-ECFP小鼠杂交以观察肠上皮隐窝的细胞。还描述了标记不同肿瘤成分（例如血管和中性粒细胞）的程序，以及将肿瘤定位为通过浆膜表面成像的程序。通过数小时的成像编辑而成的延时电影可以在肠道微环境中原位分析骨髓细胞的行为。

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