Optical imaging assisted by molecular-specific fluorescent labelling is one the main toolboxes of the life sciences. It is based on organic fluorescent dyes and suffers from the fluorescence photobleaching and intermittency, which can be circumvented by employing upconversion nanoparticles (UCNP). However, the demand of the high excitation intensity and exceptionally long photoluminescence lifetime of UCNP precludes in vivo imaging. We propose a novel image acquisition approach, where an UCNP-labelled specimen is rapidly raster-scanned, while its persistent photoluminescence signal is acquired from the entire scan area. The resultant blurred image is post-processed using a developed deconvolution algorithm to reconstruct the diffraction-limited image. The imaging of UCNP-labelled cells and UCNP tissue biodistribution demonstrated the power of the proposed approach.