Hi all, I'm working with transfected and immunostained cells (fixed) that have been imaged by confocal microscopy. In one of the cells, I observed a striking morphology — several large vacuole-like structures surrounding the nucleus. These compartments are significantly larger than typical endosomes or lysosomes, and appear to create a "storage" appearance.
I’m trying to understand what I’m looking at:
- Could these be phagosomes or autophagic vesicles?
- Is this a known stress response or a fixation/transfection artifact?
- Has anyone seen similar structures before?
I'd appreciate any input or papers that might help explain this phenotype.
Thanks in advance!
Hi, Shahar Doron
The fixation process may cause the rupture and fusion of cell membranes or organelle membranes, forming pseudovacuole structures. In addition, the accumulation or nonspecific binding of dyes may also cause visual effects similar to large vacuoles.
But you mentioned that it may be autophagic vesicles (Autophagosomes) or phagosomes (Phagosomes). These structures are usually larger than ordinary endosomes or lysosomes and are vesicle-shaped, responsible for the degradation and recycling of intracellular substances. During transfection, fixation or cell stress, autophagic activity may be enhanced, resulting in the accumulation of autophagic vesicles to form larger vacuole-like structures. If cells experience damage caused by stress or transfection, cells may initiate autophagy as a protection or cleanup mechanism. It is recommended that you co-stain with specific markers (such as LC3 or p62 to mark autophagosomes, LAMP1 to mark lysosomes) to confirm the identity of these vacuoles.
The answer to this question comes from MedChemExpress (MCE) Technical Support.
- Badges
- Science topic
- Similar topics
- Biological Science
- Anatomy
- Tissue