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Zhenfeng Zhu¹, Chris W T Leung², Xinyuan Zhao³

¹State Key Laboratory of Modern Optical Instrumentation (Zhejiang University), Centre for Optical and Electromagnetic Research, Zhejiang Provincial Key Laboratory for Sensing Technologies, JORCEP (Sino-Swedish Joint Research Center of Photonics), Zhejiang University, Hangzhou, 310058, China.

Scientific Reports

|October 16, 2015

Summary

Researchers developed photostable fluorescent probes (TPE-TPP) with aggregation-induced emission (AIE) for advanced bioimaging. This overcomes photobleaching and photodamage, enabling clearer, long-term cellular and tissue imaging.

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Area of Science:

Biomedical Engineering
Optical Imaging
Materials Science

Background:

Fluorescent probes are crucial for live-cell bioimaging but suffer from photobleaching, autofluorescence, and photodamage.
Limitations of traditional probes hinder long-term metabolic process monitoring in cells.

Purpose of the Study:

To develop a photostable fluorescent probe with aggregation-induced emission (AIE) characteristics.
To demonstrate the efficacy of the novel probe for three-photon microscopy with reduced photodamage and autofluorescence.
To compare the signal-to-noise ratio of three-photon microscopy with the developed probe against two-photon microscopy in tissue imaging.

Main Methods:

Synthesis of a novel photostable luminogen, TPE-TPP, exhibiting aggregation-induced emission (AIE).
Utilizing TPE-TPP for three-photon microscopy with femtosecond laser excitation at 1020 nm.
Evaluating probe performance in HeLa cells for photo-damage, autofluorescence, and photobleaching resistance.
Comparing three-photon microscopy (1020 nm) with two-photon microscopy (810 nm) for signal-to-noise ratio in tissue imaging.

Main Results:

TPE-TPP demonstrated excellent photostability and resistance to photobleaching under prolonged three-photon excitation.
Three-photon microscopy using TPE-TPP in HeLa cells exhibited minimal photodamage and low autofluorescence.
TPE-TPP nanoaggregates showed efficient cellular uptake and stable fluorescence.
Three-photon microscopy with TPE-TPP achieved a superior signal-to-noise ratio compared to two-photon microscopy in tissue imaging.

Conclusions:

TPE-TPP is a highly effective photostable fluorescent probe for advanced bioimaging applications.
Three-photon microscopy with TPE-TPP offers significant advantages over conventional methods, including reduced photodamage and enhanced imaging quality.
The developed AIE luminogen paves the way for improved long-term cellular monitoring and in-depth tissue imaging.