NIR-excited imaging and in vivo visualization of β-galactosidase activity using a pyranonitrile-modified upconversion nanoprobe.

β-galactosidase (β-gal) is a diagnostic biomarker of primary ovarian cancers. The development of effective fluorescent probes for investigating the activity of β-gal will be beneficial to cancer diagnosis. Herein, a near-infrared (NIR) excited ratiometric nanoprobe (DCM-β-gal-UCNPs) by assembling pyranonitrile dye (DCM-β-gal) on the surface of upconversion nanophosphors (UCNPs) was designed for the evaluation of β-gal activity in vivo. Upon the interaction with β-gal, a marked decrease of upconversion luminescence (UCL) signal in the green channel was observed owing to the luminescence resonance energy transfer from the UCNPs to pyranonitrile chromophore, whereas the NIR UCL emission at 800 nm was almost no influence. Thus, the β-gal activity could be quantitatively detected by the UCL intensity ratio of UCL/UCL with the limit of detection of 3.1 × 10 U/mL. Moreover, DCM-β-gal-UCNPs was effectively applied for monitoring β-gal fluctuation in living cells and zebrafish by a ratiometric UCL signal excited by 980 nm laser. We envision that nanoprobe DCM-β-gal-UCNPs might be used as a potential bioimaging tool to disclose more biological information of β-gal in β-gal-associated diseases in the future.