Direct Observation of Different One- and Two-Photon Fluorescent States in a Pyrrolo[3,2-]pyrrole Fluorophore.

Two-photon fluorophores are frequently employed to obtain superior spatial resolution in optical microscopy applications. To guide the rational design of these molecules, a detailed understanding of their excited-state deactivation pathways after two-photon excitation is beneficial, especially to assess the often-assumed presumption that the one- and two-photon excited-state dynamics are similar after excitation. Here, we showcase the breakdown of this assumption for one- and two-photon excitation of a centrosymmetric pyrrolo[3,2-]pyrrole chromophore by combining time-resolved fluorescence and broadband femtosecond transient absorption spectroscopy. Compared to one-photon excitation, where radiative decay dominates the photodynamics, two-photon excitation leads to dynamics arising from increased nonradiative decay pathways. These different photodynamics are manifest to different quantum yields, thus highlighting the types of time-resolved studies described here to be valuable guideposts in the design of two-photon fluorophores for imaging applications.