An Acid‐Regulated Self‐Blinking Fluorescent Probe for Resolving Whole‐Cell Lysosomes with Long‐Term Nanoscopy

Abstract

Long‐term super‐resolution imaging appears to be increasingly important for unraveling organelle dynamics at the nanoscale, but is challenging due to the need for highly photostable and environment‐sensitive fluorescent probes. Here, we report a self‐blinking fluorophore that achieved 12 nm spatial resolution and 20 ms time resolution under acidic lysosomal conditions. This fluorophore was successfully applied in super‐resolution imaging of lysosomal dynamics over 40 min. The pH dependence of the dye during blinking made the fluorophore sensitive to lysosomal pH. This probe enables simultaneous dynamic and pH recognition of all lysosomes in the entire cell at the single‐lysosome‐resolved level, which allowed us to resolve whole‐cell lysosome subpopulations based on lysosomal distribution, size, and luminal pH. We also observed a variety of lysosome movement trajectories and different types of interactions modes between lysosomes.

Homoadamantane-Fused Tetrahydroquinoxaline as a Robust Electron-Donating Unit for High-Performance Asymmetric NIR Rhodamine Development

“…Similar to classical rhodamine amide, NNR4M was present mainly as the nonfluorescent spirocyclic form at physiological pH (∼7.4). However, the fluorescence of NNR4M increased by about 2350-fold under acidic conditions reaching a maximum at pH 4 (similar to the pH in lysosomes) due to the H + -induced ring-opening process (Figure S15a). The related ring-opening mechanism can be further demonstrated by HRMS analysis of NNR4M in pH 4.0 buffer (Figure S16).…”

Visualizing Lysosomal Positioning with a Fluorescent Probe Reveals a New Synergistic Anticancer Effect

“…One study showed that lysosomes undergo two distinct types of motions during neuronal stem cell differentiation . The other study obtained and defined four representative interaction modes in lysosomes …”

“…18 The other study obtained and defined four representative interaction modes in lysosomes. 19 It has been reported that lysosomes show remarkably changed cellular distribution under altered nutrient levels. Thus, we conjectured that NIM-3 may act as a tool to investigate the dynamic response of lysosomes under different stimuli, like serum and glucose.…”

Si-rhodamine fluorescent probe for monitoring of hypochlorous acid in the brains of mice afflicted with neuroinflammation

“…According to the previous reports on the response mechanism of rhodamine probes, [27][28][29][30] an isomer of SiR-FH with a CQN bond could undergo an addition reaction with HClO. After intramolecular dehydration, the lateral N-chloro hydrazide group was cleaved and the product SiR-FH-HClO formed (Fig.…”