The study introduces OrgaPlexing, a multi-spectral organelle imaging approach, to map the interactions between six key metabolic organelles in macrophages. Using this method, the researchers found that lipid droplets (LDs) are the primary inflammatory responder organelle, forming clusters with other organelles such as the endoplasmic reticulum (ER), mitochondria, and peroxisomes. These clusters facilitate the supply of fatty acids for LD growth and support fatty acid efflux from LDs. Interfering with these multi-organellar units has direct functional consequences for inflammatory lipid mediator synthesis. The study highlights how macrophages form functional multi-organellar units to support metabolic adaptation and provides an experimental strategy to identify organelle-metabolic signaling hubs. The findings also suggest that the M–ER–P–LD unit controls inflammatory lipid trafficking and prostaglandin E2 (PGE2) production, with MIGA2 and DRP1 playing crucial roles in this process.The study introduces OrgaPlexing, a multi-spectral organelle imaging approach, to map the interactions between six key metabolic organelles in macrophages. Using this method, the researchers found that lipid droplets (LDs) are the primary inflammatory responder organelle, forming clusters with other organelles such as the endoplasmic reticulum (ER), mitochondria, and peroxisomes. These clusters facilitate the supply of fatty acids for LD growth and support fatty acid efflux from LDs. Interfering with these multi-organellar units has direct functional consequences for inflammatory lipid mediator synthesis. The study highlights how macrophages form functional multi-organellar units to support metabolic adaptation and provides an experimental strategy to identify organelle-metabolic signaling hubs. The findings also suggest that the M–ER–P–LD unit controls inflammatory lipid trafficking and prostaglandin E2 (PGE2) production, with MIGA2 and DRP1 playing crucial roles in this process.