Method for detecting activity of abdominal cavity phagocytic cells by flow cytometry

Abstract

The invention discloses a method for detecting the activity of abdominal cavity phagocytic cells by a flow cytometry, which aims at providing a method for fast, accurately, simply and easily detecting the activity of the phagocytic cells through the quantitative analysis on the number of phagocytic yeasts of the phagocytic cells on the basis of eliminating the influence of unendocytosed fluorescence yeast cells on the analysis results. The method comprises the following steps: preparing beer yeast cell freeze-dried powder; using FITC to mark the beer yeast cells; carrying out incubation on the suspension liquid of the fluorescence marked beer yeast cells and the blood serum of the mice at the room temperature; adding the suspension liquid of the abdominal cavity phagocytic cells into a hole of a culture plate; adding the fluorescence marked beer yeast cells after the incubation with the blood serum of the mice and the identical suspension liquid of the abdominal cavity phagocytic cells into the rest holes in the cell culture plate; carrying out wall adherence incubation on the cell culture plate; and using the flow cytometry to obtain data and carrying out analysis on the data. The method of the invention accurately measures the phagocytic rate and the phagocytic index, and has good repetitiveness.

Description

technical field [0001] The invention relates to the technical field of cell biology experiments, and more specifically relates to a method for detecting the activity of peritoneal phagocytes by using the fast and accurate characteristics of flow cytometry. Background technique [0002] Peritoneal phagocytes play an important role in the body's defense against the invasion of pathogens. Phagocytes are non-specific immune cells that can non-specifically phagocytize pathogens and aging diseased cells that enter the human body, participate in antigen recognition and presentation, produce various cytokines, and play an important role in the body's innate immunity. Therefore, detection of phagocyte activity is of great significance for the screening of drugs and the development of functional foods. The traditional methods for detecting the activity of phagocytes mainly include phagocytosis of chicken red blood cells and yeast. Because these methods are time-consuming, heavy workl...

AI Technical Summary

Problems solved by technology

Among them, in the method of phagocytosis of fluorescent microspheres, the particles of fluorescent microspheres are relatively uniform, and after incubation with mouse serum, immunoglobulins can be bound to the surface, which has a good detection effect, but the cost is high

Using some bacteria such as Escherichia coli to detect phagocytic activity has the following disadvantages: First, the cells are small and are easily adhered by receptors on the surface of phagocytic cells, which affects the detection results

Second, using FITC to mark these bacteria, the labeling rate is low, so certain errors will occur in the experiment

V. Miliukien reported the use of flow cytometry to detect neutrophils in peripheral blood phagocytosis of fluorescently labeled S. cerevisiae to detect its phagocytosis activity, but because it did not remove the fluorescent yeast cells that were not phagocytized, it only used forward light and side aperture Live phagocytic cells, because the individual yeast cells are larger and the number is far more than the number of granulocytes, therefore, the cells obtained by flow cytometry contain a large number of yeast cells, which reduces the number of granulocytes obtained, and at the same time mixed with A large number of yeast cells also affects the accuracy of the analysis results; moreover, in the result analysis, the number of yeast phagocytized by neutrophils was not quantitatively analyzed, so the phagocytosis index of granulocytes cannot be calculated

[0004] At present, there is no method to use fluorescently labeled yeast cells as materials to detect the activity of peritoneal phagocytes, due to V.Miliukien There are above-mentioned defects in the method, therefore, it is necessary to explore a set of more accurate, reliable and easy methods on the basis of the original detection of phagocyte activity by flow cytometry

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