Pavlova Ekaterina, Shaposhnikova Daria, Petrichuk Svetlana, Radygina Tatiana, Erokhina Maria
Faculty of Biology, Lomonosov Moscow State University, Moscow, Russian Federation.
National Medical Research Center for Children's Health, Laboratory of Experimental Immunology and Virology, Moscow, Russian Federation.
Methods Mol Biol. 2023;2635:203-215. doi: 10.1007/978-1-0716-3020-4_12.
The existing methods of quantitative analysis of phagocytosis are characterized by a number of limitations. The usual method of manually counting phagocytosed objects on photographs obtained by confocal microscopy is very labor-intensive and time-consuming. As well, the resolution of conventional flow cytometry does not allow the fluorescence detection of a large number of phagocytosis objects. Thus, there is a need to combine the rapid analysis by flow cytometry and the visualization capability by confocal microscopy. This is possible due to imaging flow cytometry. However, until now, no protocols have allowed one to quantify phagocytosis at its high intensity. The present paper presents the developed and tested algorithm for assessing the level of phagocytic activity using flow cytometry with visualization and IDEAS software.
现有的吞噬作用定量分析方法存在诸多局限性。通过共聚焦显微镜获得的照片上手动计数吞噬物体的常规方法非常耗费人力且耗时。同样，传统流式细胞术的分辨率不允许对大量吞噬作用物体进行荧光检测。因此，需要将流式细胞术的快速分析与共聚焦显微镜的可视化能力相结合。成像流式细胞术使这成为可能。然而，到目前为止，尚无方案能够在高强度下对吞噬作用进行定量。本文介绍了使用具有可视化功能的流式细胞术和IDEAS软件开发并测试的用于评估吞噬活性水平的算法。
