We report a comprehensive investigation of the photophysical properties of Hoechst 33258 (HOE) embedded in polyvinyl alcohol (PVA) films. HOE displays a bright, highly polarized, blue fluorescence emission centered at 430 nm, indicating effective immobilization within the polymer matrix of PVA. Its fluorescence quantum yield is notably high (~0.74), as determined relative to a quinine sulfate standard. In addition, we observed that HOE-doped PVA films exhibit room temperature phosphorescence (RTP) that remains visible for several seconds after UV excitation ceases. The slightly negative phosphorescence anisotropy implies that the triplet-singlet radiative transition is orthogonal to the singlet-singlet transition governing fluorescence. Notably, we observed that direct triplet-state excitation at longer wavelengths (beyond the primary absorption band) produces highly polarized RTP. We believe this possibility of direct triplet-state excitation opens new avenues for studying RTP in polymer-immobilized molecules.
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