Critical factors for flow cytometry analysis of the brain

The brain is difficult to analyze using flow cytometry due to its complex interactions with cells, high lipid content, and high autofluorescence. In this study, we investigated methods to isolate various types of brain cells with high yield and viability. The results showed that protease selection significantly affected the viability of various cell types in the brain. Differences in the developmental stage also affected cell yield and viability. Furthermore, the intensity of autofluorescence differs greatly between various regions of the brain. Additionally, we searched for neuronal indicators capable of identifying a diverse range of neurons. The ratios of various exosomes contained in neurons differ depending on the type of neuronal marker. These results revealed critical factors that must be considered when analyzing various types of brain cells using flow cytometry.