The endoplasmic reticulum (ER) is crucial for the regulation of multiple cellular processes, such as cellular responses to stress and protein synthesis, folding, and posttranslational modification. Nevertheless, monitoring ER physiological activity remains challenging due to the lack of powerful detection methods. Herein, we built a two-stage cascade recognition process to achieve dynamic visualization of ER stress in living cells based on a fluorescent carbon dot (CD) probe, which is synthesized by a facile one-pot hydrothermal method without additional modification. The fluorescent CD probe enables two-stage cascade ER recognition by first accumulating in the ER as the positively charged and lipophilic surface of the CD probe allows its fast crossing of multiple membrane barriers. Next, the CD probe can specifically anchor on the ER membrane via recognition between boronic acids and o-dihydroxy groups of mannose in the ER lumen. The two-stage cascade recognition process significantly increases the ER affinity of the CD probe, thus allowing the following evaluation of ER stress by tracking autophagy-induced mannose transfer from the ER to the cytoplasm. Thus, the boronic acid-functionalized cationic CD probe represents an attractive tool for targeted ER imaging and dynamic tracking of ER stress in living cells.

ATP and reactive oxygen species (ROS) are considered significant indicators of cell apoptosis. However, visualizing the interplay between apoptosis-related ATP and ROS is challenging. Herein, we developed a metal–organic framework (MOF)-based nanoprobe for an apoptosis assay using duplex imaging of cellular ATP and ROS. The nanoprobe was fabricated through controlled encapsulation of gold nanorods with a thin zirconium-based MOF layer, followed by modification of the ROS-responsive molecules 2-mercaptohydroquinone and 6-carboxyfluorescein-labeled ATP aptamer. The nanoprobe enables ATP and ROS visualization via fluorescence and surface-enhanced Raman spectroscopy, respectively, avoiding the mutual interference that often occurs in single-mode methods. Moreover, the dual-modal assay effectively showed dynamic imaging of ATP and ROS in cancer cells treated with various drugs, revealing their apoptosis-related pathways and interactions that differ from those under normal conditions. This study provides a method for studying the relationship between energy metabolism and redox homeostasis in cell apoptosis processes.

Nanomaterials have presented great potential for cancer therapy. However, their therapeutic efficacy is not always satisfied because of inefficient biocompatibility and targeting efficacy. Here, we report engineered extracellular vesicle (EV)-encapsuled nanoreactors for the targeting and killing of cancer cells. EVs are extracted from engineered cancer cells with surface N-glycans cut and intracellular microRNA-21 (miR-21) silenced to generate cancer-targeting membranes for the following coating of gold–polydopamine (PDA) core–shell nanoparticles. The encapsuled nanoparticles are decorated with doxorubicin (Dox), glucose oxidase (GOx), and miR-21-indicative DNA tags. Once endocytosed, the acidic pH, together with the photothermal effect of the PDA shell, can promote the release of Dox and GOx-catalyzed H2O2 generation/glucose consumption, while the DNA tags allow enhanced fluorescence imaging of miR-21 to indicate the targeting effect. The coadministration of EV-assisted delivery and cascade treatment represents a promising strategy for combination therapy.