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Near-Infrared Spontaneously Blinking Fluorophores for Live Cell Super-Resolution Imaging with Minimized Phototoxicity
Song Chen1, Jing Wang1, Daoming Guan1
1Department of Chemistry and Shanghai Key Laboratory of Molecular Catalysis and Innovative Materials, Fudan University, Shanghai 200438, China.
Analytical Chemistry
|June 18, 2024
Summary
Researchers developed near-infrared (NIR) blinking fluorophores for super-resolution microscopy. These probes enable high-resolution imaging of living cells with reduced light toxicity, advancing cellular ultrastructure studies.
Area of Science:
- Biophysics
- Cellular Imaging
- Organic Chemistry
Background:
- Single-molecule localization microscopy (SMLM) requires high laser power, causing cellular toxicity and limiting live-cell imaging.
- Existing fluorophores often suffer from photobleaching and limited emission under physiological conditions.
Purpose of the Study:
- To develop novel near-infrared (NIR) spontaneously blinking fluorophores for SMLM.
- To enable high-resolution live-cell imaging with reduced phototoxicity.
Main Methods:
- Synthesized NIR fluorophores by combining rhodamine spirolactams and merocyanine derivatives.
- Characterized fluorophore properties including blinking, emission rate, and duty cycle.
- Performed live-cell imaging experiments using time-lapse SMLM with NIR illumination.
Main Results:
- Developed NIR fluorophores with a low duty cycle (0.18%) and high emission rate (26,700 photons/s) at 3.93 kW/cm2.
- Demonstrated reduced cellular toxicity under NIR illumination (721 nm) compared to visible light.
- Achieved time-lapse super-resolution tracking of mitochondria (69.4 nm resolution) and ER ultrastructure in living cells.
Conclusions:
- NIR spontaneously blinking fluorophores are effective for live-cell SMLM.
- This technology minimizes phototoxicity, allowing for prolonged observation of cellular dynamics.
- Enables advanced studies of cellular ultrastructure and function in living systems.
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