Chemical modulation of fluorescence lifetime of fluoroboron dipyrrole dyes and application in multi fluorescence lifetime imaging

Reprinted: X-MOL

Fluorescence lifetime imaging (FLIM) technology has become an important tool in chemical biology research. Compared with traditional imaging methods based on fluorescence intensity, fluorescence lifetime adds an additional imaging channel on the basis of fluorescence intensity. By applying fluorescent probes with different fluorescence lifetimes and similar spectral properties (excitation and emission wavelengths), multiple targets can be imaged and split simultaneously within the same spectral channel. Recently, the team led by Zhang Xin from the School of Science at Xihu University successfully achieved systematic modulation of the fluorescence lifetime of fluoroboron dipyrrole dyes in green and red spectral channels through chemical regulation of photo induced electron transfer (PET) and intramolecular charge transfer (ICT) processes.

The existing multiple fluorescence lifetime imaging is mainly achieved by selecting different types of probe molecules, and the number of imaging channels is limited by the types of these molecules within the same spectral channel and the small range of fluorescence lifetime changes. Regulating the fluorescence lifetime of molecules of the same type and obtaining a series of molecular libraries with different fluorescence lifetimes can greatly expand the number of channels for multiple fluorescence lifetime imaging. However, so far, there is still a lack of systematic research on regulating the fluorescence lifetime of fluorescent small molecules.

Reprinted: X-MOL

Fluorescence lifetime imaging (FLIM) technology has become an important tool in chemical biology research. Compared with traditional imaging methods based on fluorescence intensity, fluorescence lifetime adds an additional imaging channel on the basis of fluorescence intensity. By applying fluorescent probes with different fluorescence lifetimes and similar spectral properties (excitation and emission wavelengths), multiple targets can be imaged and split simultaneously within the same spectral channel. Recently, the team led by Zhang Xin from the School of Science at Xihu University successfully achieved systematic modulation of the fluorescence lifetime of fluoroboron dipyrrole dyes in green and red spectral channels through chemical regulation of photo induced electron transfer (PET) and intramolecular charge transfer (ICT) processes.

The existing multiple fluorescence lifetime imaging is mainly achieved by selecting different types of probe molecules, and the number of imaging channels is limited by the types of these molecules within the same spectral channel and the small range of fluorescence lifetime changes. Regulating the fluorescence lifetime of molecules of the same type and obtaining a series of molecular libraries with different fluorescence lifetimes can greatly expand the number of channels for multiple fluorescence lifetime imaging. However, so far, there is still a lack of systematic research on regulating the fluorescence lifetime of fluorescent small molecules.

Figure 2. Multi fluorescence lifetime imaging targeting different proteins

The author subsequently validated its application in multi-channel fluorescence lifetime imaging in vitro and in vivo. The author linked molecules with different fluorescence lifetimes to elastin like peptides (ELPs) that can undergo liquid-liquid phase separation. After forming droplets, the droplets with different fluorescence lifetimes can be clearly separated and displayed through FLIM. Using the same strategy, probe molecules connected to self labeled protein tags can be accurately separated by FLIM after labeling nuclear phosphoprotein NPM1 and histone H2B separately. In the in vivo experiment, using probes with different fluorescence lifetimes connected to targeted endoplasmic reticulum and lysosome groups, the authors applied FLIM to achieve simultaneous imaging of endoplasmic reticulum and lysosomes in U-2 OS cells in red and green channels.

This study provides an effective method for systematically regulating the fluorescence lifetime of BODIPY dyes, establishing a molecular library in two spectral channels that can be applied to in vitro and in vivo multiple fluorescence lifetime imaging, and similar strategies may be applicable to the fluorescence lifetime regulation of other types of fluorescent groups.

This achievement was recently published in Angew On Chem. Int. Ed., the first authors of the article are Ma Junbao, a postdoctoral fellow at West Lake University, and Luo Feng, a doctoral student.

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