Lipid droplets (LDs) are essential organelles responsible for storing neutral lipids and maintaining lipid homeostasis in cells. Dysregulation of LD degradation is closely associated with various metabolic disorders and neurodegenerative diseases. Lipophagy, a selective autophagy pathway, mediates the transport of LDs to vacuoles or lysosomes for degradation; however, the mechanisms underlying selective recognition and recruitment remain unclear. In this study, Saccharomyces cerevisiae was employed as a model organism to systematically screen 48 LD-associated membrane protein deletion strains using high-throughput fluorescence microscopy, aiming to identify key factors involved in LD autophagy. Our results demonstrate that LDs are degraded via autophagic pathways involving both microautophagy and macroautophagy. We further established optimal conditions for the induction of LD autophagy and identified 13 gene deletions that markedly impair this process, suggesting their potential roles as receptors or regulators of lipophagy. This work advances our understanding of the molecular mechanisms governing LD autophagy and its contribution to cellular lipid homeostasis, providing a conceptual framework for developing therapeutic strategies for metabolic and neurodegenerative disorders.