Deciphering interior polarity of lysosome in live cancer and normal cells using spectral scanning microscopy

A lysosome specific, pH tolerant, and polarity-sensitive fluorescent probe (LyPol) is designed and synthesized for the determination of lysosomal polarity in live cells. LyPol possesses an intramolecular charge transfer (ICT) properties with high quantum yield in water and in other polar solvents such as methanol, ethanol, dimethyl sulfoxide, acetonitrile, etc. The fluorescence maxima and lifetime increase linearly with a non-specific manner with an increase in the polarity of its surrounding environment. A morpholine group connected with an alkyl linker acts as a lysosome directing moiety, which is attached to the fluorescent core of LyPol. The selective localization of LyPol inside the lysosome was confirmed with live-cell confocal imaging. Further, the spectral scanning confocal technique was utilized to determine the emission spectrum of LyPol inside lysosome, and the polarity turns out to be quite lower as compared to water. Moreover, the combined spectroscopic and live-cell microscopy confirms that the interior of the lysosome is significantly non-polar in cancer cells compared to normal cells. We believe that this report on the measuring polarity inside the biological system with a solvatofluorochromic probe will be of immense interest to researchers working in the multidisciplinary area of biophysics, microscopy, chemical biology, and organelle biology.