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- W4313403537 abstract "Abstract Although engulfment is a hallmark of microglia function, fully validated platforms that facilitate high-throughput quantification of this process are lacking. We therefore developed FEAST ( Flow cytometric Engulfment Assay for Synaptic Terminals ) to enable interrogation of in vivo engulfment of synaptic material by brain resident macrophages at single-cell resolution. We optimized FEAST for two different analyses: quantification of fluorescent material inside live cells and of engulfed endogenous proteins within fixed cells. To overcome false-positive engulfment signal, we introduced an approach suitable for interrogating engulfment in microglia from perfusion-fixed tissue. As a proof-of-concept for the specificity and versatility of FEAST, we examined the engulfment of synaptic proteins after optic nerve crush as well as myelin in two models of demyelination and found that microglia, but not other brain-associated macrophages, engulf in these contexts. Our work underscores how FEAST can be utilized to gain critical insight into functional neuro-immune interactions that shape development, homeostasis, and disease." @default.
- W4313403537 created "2023-01-06" @default.
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- W4313403537 date "2022-12-19" @default.
- W4313403537 modified "2023-10-18" @default.
- W4313403537 title "Interrogating microglial engulfment of synaptic terminals and myelin using FEAST" @default.
- W4313403537 doi "https://doi.org/10.21203/rs.3.rs-2363013/v1" @default.
- W4313403537 hasPublicationYear "2022" @default.
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