Data for "Ultrafast in vivo spectroscopy reveals photosynthetic energy and charge transfer under physiological conditions"

Ultrafast spectroscopy has transformed our understanding of photosynthesis, yet most insights have come from in vitro studies outside of the native cellular environment, leaving open the question of whether the same mechanisms operate in vivo. Here, we address longstanding challenges of in vivo ultrafast measurements and demonstrate that femtosecond transient absorption and two-dimensional electronic spectroscopy can directly resolve photosynthetic energy and charge transfer in living cells of the cyanobacterium Synechocystis sp. PCC 6803 under physiological, open reaction centre conditions. We resolve three key steps of photosynthesis: energy transfer from bulk antenna chlorophylls to reaction centres, energy transfer from phycobilisomes to both photosystems I and II, and signatures of electron transport from photosystem II to photosystem I. The agreement between our results and those from isolated-complex lifetimes reinforces decades of mechanistic work. However, the in vivo framework captures the additional complexity and competing pathways of native membranes. Our results establish a platform for investigating photosynthetic regulation and adaptation at the cellular level, and provide quantitative benchmarks for the engineering of artificial and biohybrid light-harvesting systems inspired by the efficiency and robustness of nature. Data files for this paper: Files labelled as Map_2DES correspond to the data presented in Figure 2 Files labelled as LifetimeDensityMap correspond to the data presented in Figure 3 File labelled 610 nm pump corresponds to the data presented in Figure 4 Files laballed 680 nm pump corresponds to the data presented in Figure 5 Text file contains code written for the lifetime density map analysis