Investigation of flow cytometry for analysis of mouse brain

The brain is difficult to analyze using flow cytometry because of its complex interactions of cells, high lipid content, and high autofluorescence. In this study, we investigated methods to isolate various types of brain cells with high yields and viability. The results showed that proteinase selection had a significant effect on the viability of various types of cells in the brain. The differences in the developmental stage also affected cell yield and viability. Moreover, the intensity of autofluorescence was found to differ greatly among the various regions of the brain. We also searched for neuronal markers that recognize a wide range of neurons. The ratios of the various exosomes contained by neurons differed depending on the type of neuronal marker. We also developed a method to quantify microglial phagocytosis in large quantities and in a short time using flow cytometry. These results have revealed critical factors that must be considered when analyzing various types of brain cells using flow cytometry. Overall design: Treat mouse telencephalon with papain at P8, centrifuge at 24% SIP, and identify precipitated cell types.