The photophysical properties of a heavy atom-free BODIPY derivative with a twisted π-conjugated framework were studied. Efficient intersystem crossing (ISC quantum yield: 56%) and an exceptionally long-lived triplet state were observed (4.5 ms in solid polymer film matrix and 197.5 μs in solution). Time-resolved electron paramagnetic resonance (TREPR) spectroscopy and DFT computations confirmed the delocalization of the triplet state on the whole twisted π-conjugated framework and the zero-field-splitting (ZFS) D parameter of D = -69.5 mT, which is smaller than that of 2,6-diiodoBODIPY (D = -104.6 mT). The electron spin polarization (ESP) phase pattern of the triplet state TREPR spectrum of the twisted BODIPY is (a, a, e, a, e, e), which is different from that of 2,6-diiodo BODIPY (e, e, e, a, a, a), indicating that the electron spin selectivity of the ISC of the twisted structure is different from that of the spin orbital coupling effect. According to the computed spin-orbit coupling matrix elements (0.154-1.964 cm-1), together with the matched energy of the S1/Tn states, ISC was proposed to occur via S1→T2/T3. The computational results were consistent with TREPR results on the electron spin selectivity (the overpopulation of the TY sublevel of the T1 state). The advantage of the long-lived triplet state of the twisted BODIPY was demonstrated by its efficient singlet oxygen (1O2) photosensitizing (ΦΔ = 50.0%) even under a severe hypoxia atmosphere (pO2 = 0.2%, v/v). A high light toxicity (EC50 = 1.0 μM) and low dark toxicity (EC50 = 78.5 μM) were observed for the twisted BODIPY, and thus the cellular studies demonstrate its potential as a novel potent heavy atom-free photodynamic therapy (PDT) agent.