Apoptotic vesicles (ApoVs) are membrane structures formed during cell apoptosis and play crucial roles in homeostasis maintenance, signal transduction, and immune regulation. Importantly, ApoVs inherit the properties and contents of parental cells that show great potential in the diagnosis and treatment of diseases. Monitoring the formation process of ApoVs (such as quantity, morphological changes, release rules, etc.) can reveal the regulatory mechanism of apoptosis, and is also helpful for optimizing the preparation and application of ApoVs. However, due to the limitations of existing technologies, the formation processes of ApoVs have been challenging to precisely and entirely capture. Herein, we subtly constructed a versatile AIEgen (ADTP) that could induce ApoVs production and in situ monitor the formation process, and it was successfully applied to explore the formation mechanism of ApoVs. ADTP specifically targeted the plasma membrane, and it could effectively induce apoptosis under laser irradiation, so it was able to dynamically monitor the entire formation process of ApoVs and had validated ApoVs formation from membrane protrusions (including filopodia, tunneling nanotubes, and retraction fibers). Further investigation revealed that ApoVs derived from membrane protrusions with different components exhibited significant heterogeneity. Additionally, the near-infrared emission characteristic of ADTP was compatible with the stimulated emission depletion (STED) microscopy equipped with a 775 nm depletion laser, enabling high-resolution visualization of detailed dynamic changes in membrane protrusions during ApoVs formation. This work provided powerful tools for tracking the entire ApoVs formation process and also offered crucial scientific evidence for revealing the ApoVs formation mechanism.
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