This article reviews the state of the art of ultrafast transient absorption microscopy, discusses current experimental concepts and highlights future challenges. The advantages of transient absorption microscopy over other micro-spectroscopic techniques are its high optical resolution combined with high temporal resolution as well as its ability to study non-fluorescent and weakly fluorescent molecular species and to probe excited-state processes. In conventional transient absorption spectroscopy the spectroscopic information usually presents a spatial average over the focal spot of the typically weakly focused probe beam. Transient absorption microscopy, however, enables investigations of the excited state dynamics in individual microscopic areas of a sample. Hence, the technique does not only yield detailed morphological information based on a label-free molecular contrast, but also gives insight into the ultrafast morphology-dependent photoinduced processes in heterogeneous samples. Different variations of transient absorption microscopy have found a number of applications ranging from material sciences to biology, which are discussed in this review together with different setup modifications and approaches towards transient absorption spectroscopy with spatial resolution below the diffraction limit.

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An in cellulo study of the ultrafast excited state processes in the paradigm molecular light switch [Ru(bpy)2dppz]2+ by localized pump-probe spectroscopy is reported for the first time. The localization of [Ru(bpy)2dppz]2+ in HepG2 cells is verified by emission microscopy and the characteristic photoinduced picosecond dynamics of the molecular light switch is observed in cellulo. The observation of the typical phosphorescence stemming from a 3MLCT state suggests that the [Ru(bpy)2dppz]2+ complex intercalates with the DNA in the nucleus. The results presented for this benchmark coordination compound reveal the necessity to study the photoinduced processes in coordination compounds for intracellular use, e.g. as sensors or as photodrugs, in the actual biological target environment in order to derive a detailed molecular mechanistic understanding of the excited-state properties of the systems in the actual biological target environment.

Two‐photon laser writing is used here to fabricate 3D proteinaceous microstructures with photothermal functionality in the near‐infrared spectral region and tunable elasticity. The photo‐cross‐linking is initiated in bovine serum albumin (BSA) by rose bengal or methylene blue and the photo‐thermal effect arises from gold non‐spherically symmetric nanoparticles dispersed in the ink. Massive energy transfer of the plasmonic resonances of the gold nanoparticles to methylene blue prevents effective photo‐crosslinking of BSA. However, stable microstructures with photo‐thermal functionality can be fabricated in the rose bengal proteinaceous inks. On these microstructures, with a gold atom concentration as low as 1% w/w, a highly localized temperature increase can be quickly (≅1 s) reached and maintained under continuous wave laser irradiation at 800 nm. The photothermal efficiency under continuous wave laser irradiation depends on the thickness of the microstructure and can reach 12.2 ± 0.4 °C W−1 These proteinaceous microstructures represent therefore a promising platform for future applications in the fields like physical stimulation of cells for regenerative nanomedicine.