FEAST: A flow cytometry-based toolkit for interrogating microglial engulfment of synaptic and myelin proteins.

ABSTRACT: Although engulfment is a hallmark of microglia function, fully validated platforms that facilitate high-throughput quantification of this process are lacking. Here, we present FEAST (Flow cytometric Engulfment Assay for Specific Target proteins), which enables interrogation of in vivo engulfment of synaptic material by brain resident macrophages at single-cell resolution. We optimize 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 signals, we introduce 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 examine the engulfment of synaptic proteins

SUBMITTER: Dissing-Olesen L 

PROVIDER: S-EPMC10533836 | biostudies-literature | 2023 Sep

REPOSITORIES: biostudies-literature