Abstract：
Phagocytosis is an essential innate immunity function in humans and animals. A decrease in the ability to phagocytize is associated with many diseases and aging of the immune system. Assessment of phagocytosis dynamics requires quantification of bacteria inside and outside the phagocyte. Although flow cytometry is the most common method for assessing phagocytosis, it does not include visualization and direct quantification of location of bacteria. Here, we used double-labeled Escherichia coli cells to evaluate phagocytosis by flow cytometry (cell sorting) and confocal microscopy, as well as employed image cytometry to provide high-throughput quantitative and spatial recognition of the double-labeled E. coli associated with the phagocytes. Retention of pathogens on the surface of myeloid and lymphoid cells without their internalization was suggested to be an auxiliary function of innate immunity in the fight against infections. The developed method of bacterial labeling significantly increased the accuracy of spatial and quantitative measurement of phagocytosis in whole blood and can be recommended as a tool for phagocytosis assessment by image cytometry.
摘要：
吞噬作用是人类和动物中必不可少的先天免疫功能。吞噬能力的降低与许多疾病和免疫系统的老化有关。吞噬作用动力学的评估需要对吞噬细胞内部和外部的细菌进行定量。尽管流式细胞术是评估吞噬作用的最常用方法，它不包括细菌位置的可视化和直接量化。这里,我们使用双标记的大肠杆菌细胞通过流式细胞术（细胞分选）和共聚焦显微镜来评估吞噬作用，以及采用图像细胞计数来提供与吞噬细胞相关的双标记大肠杆菌的高通量定量和空间识别。病原体保留在骨髓和淋巴细胞表面而不内化被认为是先天免疫在抵抗感染中的辅助功能。开发的细菌标记方法显着提高了全血中吞噬作用的空间和定量测量的准确性，可以推荐作为通过图像细胞计数进行吞噬作用评估的工具。
