The first post-pandemic face-to-face international conference in Biophotonics took place in Porto in April 2022. High quality and inspiring presentations have been delivered by leading scientists and internationally recognized professionals, standing on the forefront of Biophotonics research and representing broadness of the field. A selected number of these presentations resulted in the papers that are presented in this special issue. The cover images were selected from Hoffer et al., Lomanovskaya et al., Li et al., and Burton et al. (clockwise).