Dark-vacuole Bodies Studying by High-resolution Label-free Microscopy Assisted with Fluorescence Technology

The intracellular logistics system, consisting of vesicles, plays a crucial role in cellular transport. However, there is a lack of research on the types and functions of intracellular vesicles, and new technologies are needed for further investigation. Recently, researchers have discovered a new subcellular structure known as Dark-vacuole bodies. The composition, function, and potential synergy with other organelles of these Dark-vacuole bodies remain unclear. In this study, we utilized the high-resolution label-free Fourier ptychographic microscopy, developed by our research group, along with fluorescence confocal technology, to study and analyze Dark-vacuole bodies. Our findings provide evidence of the influence of Dark-vacuole bodies on the morphology, distribution, movement, and cell cycle of living cells. Specifically, we analyzed the effects of drug induced stimulation of lipid drops and endosomes, promotion of cell endocytosis, and induction of cellular senescence on Dark-vacuole bodies. Our results indicate that Dark-vacuole bodies show little correlation with lipid drops and endocytosis vesicles, but are significantly associated with late endosomes. Furthermore, cellular senescence leads to a significant increase in the number and size of Dark-vacuole bodies. This study serves as a foundation for further confirming the nature of Dark-vacuole bodies as new organelles.